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So it's a pity to find ourselves in the last session of what's been a very stimulating day.

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Yes, I mean, it's a witness to the impact it's had.

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We've seen how wide the ramifications of what's come out of this lab.

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Today's event celebrating an entire career and it's a unique stage that afflicts all of us.

5
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Sooner or later, try as we may to delay it.

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This day of reckoning cannot be put put off indefinitely. And at some point,

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we all have to acknowledge that we have been privileged with a unique opportunity to contribute

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in a small but significant way to explaining and understanding the world we live in.

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I suspect that the difficulties we all have in making this transition are exacerbated by the suspicion,

10
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if not fear, that we may not have fully realised our potential.

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There are more discoveries that remain out there and remain elusive. If only we had a little longer.

12
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A second difficulty, of course, is the discovery process is such fun.

13
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Essentially, it's addictive, and it's very hard to give that up.

14
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Now, though, I'm sure that Dave may also have some regrets hanging up his boots in this regard.

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I'm confident that these will not be compounded by regrets about his achievements.

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Because these have been numerous and more important have had far reaching ramifications.

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They've. Your career has been one that all of us would be envious of.

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I sometimes we think of envy as being a bad thing, but I think sometimes it's actually very important to be envious because it means

19
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there's something you wish you had and with your career we would all be envious of.

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Moreover, as witnessed by the speakers at this symposium, your career has spawned numerous others,

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creating intellectual and scientific waves that continue to ripple around the world.

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This symposium has documented many of these,

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and we should all be hugely grateful to Lydia and those who have helped her who have made this symposium possible,

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Lydia, thank you very, very much for.

25
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Lastly, I'd like to thank all of you who have braved the inconvenience of international travel during the COVID era to attend,

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as well as contribute to this wonderful celebration. Now, it's important to remember that in addition to being a magnificent molecular detective,

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Dave has been a wonderful colleague to all of us here in biochemistry. I'm sure to all of his colleagues in Glasgow and in Sussex previously.

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And he's been an invaluable mentor for for, for innumerable students, postdocs, young faculty.

29
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And I think we in biochemistry are delighted that actually he's not going to disappear at all.

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In fact, I suspect he's going to be even more engaged than ever because he's not having to run his group.

31
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He's going to be here asking questions and helping people and asking difficult questions,

32
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and [INAUDIBLE] continue, I'm sure, to be an active participant in the intellectual millier of this department.

33
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I just would like to quote, and I'm going to then say a tiny little bit about Dave coming here to Oxford and what other things he's done here,

34
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but I just want to end with a quote from Francois Jacob, who I think probably inspired most of us in this room.

35
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I mean, really, this is an answer to Neil's question what's wrong with maths?

36
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And Francois Jacob had, I think, a pretty good answer to this.

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Now some of you may agree,

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and the wonderful thing is you can agree and disagree about it because he actually said something that one could disagree with.

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He said the task of a scientist is is not to explain the world in terms of what we already know, as is done in the field of mathematics.

40
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But in terms of what can be imagined. And what he meant there is that the task the son to the real driving force of

41
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science and this is something that Dave has been absolutely brilliant doing,

42
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is to understand enough to realise that actually what you're now studying cannot be explained about what is already out there.

43
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That is the Cartesian. They've got it completely wrong.

44
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You've got to say you cannot explain it in terms of this body of knowledge we built up and you have to imagine new forces to do so.

45
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And and then the process and imagining those new forces is that magical initially and eventually,

46
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then you have to integrate them into the existing knowledge.

47
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And so I think in a way, if Neil's question what is wrong with maths, I think Jacob was saying it is actually a different process.

48
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Maths is a very important part of this whole process.

49
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The fundamentally different parts of it and Dave's career is a wonderful example of that of probing and probing, trying to explain.

50
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But but the key thing is realising you can't and and I've seen witnessed him doing this and day in and day out,

51
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week in, week out, month in, month out. I think it's actually not unique to science.

52
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I'm sure Russell knows exactly what I'm talking about. And so, Dave, thank you very much for forcing us to imagine new things.

53
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Dave, as many of you heard, was came here in ninety four.

54
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He was. He came here and replaced Paul Paul, nurse and the chair of microbiology.

55
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It's interesting that sort of they've moved, you know, there was a back and forth between Scotland and Sussex,

56
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and Dave went from Sussex to Scotland and Paul went from Sussex to to and from from from from Scotland to Sussex.

57
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And I think that one of the things that you know, when he moved here, he he he continued,

58
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what I would call this virtuous circle of genetics and biochemistry.

59
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You use genetics to find stuff. You then study it biochemically.

60
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But of course, you don't know whether that's real or not, and you have to go back and do biochemistry and you create this virtuous circle.

61
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And I think this is a philosophy that, you know, genetics now seems to sort of rule the roost, and geneticists seem to think that,

62
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you know, you, you know, you do the genetics and that's the end of the story, but you have to go round the circle.

63
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And I think I think this is something that modern genetics has yet to to to fully appreciate.

64
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I mean, it is it is very, very powerful to human genetics is far more powerful than yeast or ecology ever was.

65
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But one has to realise that in order to use this information, you've got to go out and do the biochemistry and then go back and do more genetics.

66
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And of course, you know, one of the key developments was doing was was was coming to to Oxford.

67
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They've obviously started studying the dynamics of chromosomes and the proteins that determining and then and so reinvented his career,

68
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moving on from genetics and biochemistry to to to the use of state of the art imaging.

69
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And that's another remarkable thing is how he's managed to go on reinventing himself in that manner.

70
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But he didn't just do that because he also had an enormous role in choosing Hawkins Brown as

71
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architects for this building and helping to get this building off the ground together with Jonathan.

72
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Of course, they were the sort of managers, the scientific managers he kept Russell under control.

73
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He was also very important to prevent Russell turning this into a glass menagerie.

74
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She threatened to do at one point it would have ended up more like a crick where you can't hear yourself think.

75
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And so the one of the remarkable things of this building, not only is it a wonderful place to live,

76
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but you actually actually hear what other people are saying, which is very important.

77
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And Dave was very important in pinpointing potential flaws in the acoustics, which actually were fixed.

78
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So I think we'll start then with with Russell and Dave.

79
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Thank you very much for everything. So thank you very much.

80
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There's going to be an abrupt, if you imagine, sort of handbrake turn on the intellectual side at this stage.

81
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So, so careful. Yeah. I'm [INAUDIBLE] terrified anyway.

82
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Yes. So I operate or as operate very much as a team like you do, but at a completely different scale.

83
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And so therefore, I'm very much less with much less precision.

84
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It's got to be said. And as you can probably guess and and so it isn't.

85
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I mean, it's quite amazing how you talk about things which you know, are not visible to the human eye and how you're so confident about these,

86
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these sort of three dimensional things, which are obviously very, very microscopic levels.

87
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So ideally rather brutal, ill formed, imprecise things together that you can switch the lights on and off or bang your head against.

88
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I think the other thing I should say is that I will be speaking in English or trying to speak in English,

89
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whereas most as though it's a very much enjoyed has left me very much behind.

90
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In some of the discussions about.

91
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I was asked a couple of years I could get that, but yeah, I did win the chemistry prise, but that's bloody useless here.

92
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That's I can tell you. Yeah. So I think the first thing to say is that I am not the architect.

93
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I am a member of the team. So this is more and more my wife's here.

94
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And also it's people like Oliver and Louisa are very much positive, so I shouldn't take all the credit,

95
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although I do most of this standing up and talking about it, so I'm going to go forward.

96
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So there you go. So really the point to make is we can say splendid pictures of ourselves.

97
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But David is part of a massive organisation, sometimes to his regret and and I'm part of I'm the maverick in the organisation.

98
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Maybe we both are. But that's just to talk about how I think the other thing to say is right, that's just good.

99
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Background quickly is it's taken about 15 16 years so far.

100
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I think that's about right. So it took probably about five years to get to the end of phase one.

101
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And then there's a big, long gap and then we obviously phase two is now being moved into.

102
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So I was going to say that architecture takes quite some time, but clearly science takes quite a long time, too.

103
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So in parallel on that, I think the next thing to say is, unfortunately, when the project starts, it is not a blank sheet of paper.

104
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Or sometimes we wish it is, and sometimes people imagine it is, and they can have whatever they like in the world.

105
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But of course, there are many constrictions and these were some of them.

106
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So I mean, maybe this is a heresy, but we did actually knock down all these buildings to build this,

107
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and that was pretty contentious at the time and quite a nice way.

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For instance, a couple of the buildings have actually been funded by people or people's families,

109
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and so we had to actually ask the right to take the building down.

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And I think that my favourite was the little Donalds Woods where I used to go and see David in his lab,

111
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where he'd seen as I tried to bang his head against the ceiling, but he seemed to love it.

112
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Another the hands crab. And I don't know if this.

113
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If you haven't been here a lot as built in two phases and the first phase is up to

114
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the sort of lifts and actually the hands credit tower stayed up for quite a while.

115
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And then even when it came down, unfortunately, it became a car park.

116
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Now that was a bad move because the people are more in love with cars than they are in architecture and science.

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And so actually it was the site for like a rock and then it was a car park and that

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slowed up the whole process for quite a while until the second phase could be done.

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I think the second thing that I wanted to talk about and it's really the parallels I should say I don't envy you in the slightest that isn't,

120
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but is the parallels between architecture and science and thinking about this,

121
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this thing today, I very we did a very extensive art programme for the first phase and then the second phase, which is really pretty unique.

122
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What was great about it is, I think once the department got the idea that other departments had done a bit of art, it became a sort of arms race.

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So we got very competitive about how much art they could get into the building.

124
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But one of the artists was a bloke who Tim had had shortlisted for the Turner prise in one two years before,

125
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and he worked with David Samson, and they were both very interested in the digital arts.

126
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And so they learnt this sort of parallels between art and science. And I think quite interesting today,

127
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that sort of parallels between architecture or designing buildings is probably more down to earth phase and science in that.

128
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It isn't something you just get started, you have to design this sort of process.

129
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You do your research. You you go to the site, you look at the processes within, you talk to people,

130
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you find out what's been done before and then you design this process and you look about what you're trying to achieve.

131
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And I think quite often you've heard today about the questions, what's the question?

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What's the question? I think that's an individual thing. What what is the point in this building?

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What's the question it's asking? And I think the question we're asking here is, can we have a building one that brings brought a whole lot of demands,

134
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but a whole load of people together very closely? And would that create better science, I think, is a side issue.

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If we put them in a half decent building that wasn't leaking or, you know, I mean,

136
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I'm against the whole is this sort of leaky fridges and taped over windows.

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If we actually made it a nicer place to be or a half reasonable place to be?

138
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Would that change the quality of their lives?

139
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So I think that's the question, and maybe you can tell me whether we've won or not, whether whether we've passed the question.

140
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And so then onto the next slide. But.

141
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So then I think the other thing that was quite interesting, and again,

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and there's this sort of just going back, one of the things we did is we do building visits.

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So what are the great things that we bring in the wonderful when there's a there's a man called Dennis O'Driscoll who isn't here,

144
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that's a bit of a shame that he's the he's the sort of fixer behind this sort of academic.

145
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He is the best account of his role is now, but he was definitely the fix man who helped with the money.

146
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And one of the things that he helped organise with us was building visits.

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So with David and Jonathan, we went fact.

148
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We went to see camp in Vienna. And so we travelled less glamorous.

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I should say we went to Dundee and Birmingham.

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Vienna was a bit of a hard place, but we went to look at other people's laboratories and how they were working and how they were laid out.

151
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But on top of that, I think it was quite a lot about the actual functions,

152
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about how deliveries and we started to learn about how much stuff comes in and goes out every day and how some of it is poisonous or some

153
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of which is radioactive and how important it is to bring that in and out and how how much backstage stuff has to be in sovereign state.

154
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One of you is talking very nicely. I think about your technician, the technician lady came up.

155
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That's right. And how important they are that it's not, you know, not some sort of magician that just appears on stage.

156
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There's a whole backup behind you. So it was learning about that and the sort of front of house and back of house.

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And then because it's Oxford and I'm sure it's anyway, you've been told you're there was gossip,

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fear of change and hopes and dreams and a lot of politicking that we were very much helped.

159
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And I think obviously we dealt with Kim and David and Jonathan at high level, but there was politicking at levels and vice chancellors that we cannot.

160
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Yeah, we got to hear about why that sort of gossip.

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And then I think there's some wonderful hand-holding by canals to David and his team and also for them to us and a lot of talking,

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some of it with alcohol. And this is a this book.

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I think we publish this book about the whole process.

164
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One of the interesting things is probably a particularly good example of that,

165
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that we noticed that David had actually built a house this big conversion and that he was quite so famous.

166
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And how much you talk about things in three dimensions, how you are modelling and the how some symbol of the drawings,

167
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somehow we were talking earlier that maybe these hand sketches sometimes have more about them than the very

168
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sophisticated computer that they sense that it's a three dimensional thing and you were bringing that to the building.

169
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I think the other thing is that this is this is hand-built.

170
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It just shows you how it is that, but that we did actually go through the process in this really for data and asking,

171
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you know what chemicals and gases are going to be there, how they what lab equipment and it was and it was mopped up.

172
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In fact, there was a mock up for that in the Hans Krabbe building and everybody used it.

173
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What was really nice, though, is it began to do this. What that, you know,

174
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very expensive sort of consultants would tell you about is it's about change management

175
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that actually this whole process of talking was about bringing the departments together.

176
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So actually going to the data, visiting other buildings, building models, measuring how far it back to back bench that,

177
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doing that and then spreading the word is is a bit of change control in itself.

178
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They get. I mean, this is a bit I'm excited, but it's probably hard anybody else is nervous.

179
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But it's a wonderful process to hear we are going to the factories where they're making the glass and then bringing them to site.

180
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And that although this is less of a hands on process than it used to be, we have to talk about like a real world, you know, oppressed around that.

181
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So how it not?

182
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It's usual it's back, so they're sort of billed as a very central mess, and again, there was visiting the building from the team as we went along.

183
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And then back to a couple of things.

184
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One of the things we forget at the time there was protests across the road about vivisection and about research with animals.

185
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And it was quite shocking at the time, but wonderful. And we have supported very much debate to be proud of science.

186
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And it's a wonderful that you walk around the building,

187
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you look and you see people actually working to be transparent, proud of science, not being hidden away.

188
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And I think that's wonderful. That's so that says, people walk around undergraduates and graduates and looking in and say, this is going to be great.

189
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And then also about trying to challenge the silo mentality and the cafe.

190
00:21:12,550 --> 00:21:23,650
And Dave is always very keen that it must be a cafe that people get together and these people are working hard at their science and they got for data.

191
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I love this one because nobody in it has got their lab coats, and so we did really go into how things function to the absolute ends degree.

192
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And I'm sure maybe that being outdated rather quickly couple both on the art.

193
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And this is so we brought the art to site.

194
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Nicky Hirst And again, this whole new thing about art become part of it, but actually being really integrated with the science and science,

195
00:21:50,290 --> 00:21:54,880
talking to art and thinking about how you represent your scientific work.

196
00:21:54,880 --> 00:22:00,280
And is that or is that computer science? And then the last couple of sides.

197
00:22:00,280 --> 00:22:04,500
So here we are. You're probably still learning.

198
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To work with your new piece of equipment, a new piece of kit, and then I think we get this far and then you had to do it all over again.

199
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What happens is that over again, having gotten used to being in your building or at least half of it, that it had to happen all over again.

200
00:22:22,190 --> 00:22:26,780
And there is I think that it's not a CGI that really is how it is.

201
00:22:26,780 --> 00:22:32,180
And then you go back to the number of people, I think Raymond Drax in there, Dennis is in the top.

202
00:22:32,180 --> 00:22:38,000
And this is about for a 50th of the number of people it took to build the building.

203
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There you go. I think that's me. There you go. Also, thank you very much for coming.

204
00:22:48,920 --> 00:22:54,960
I think it's very, very noble of you to sit through all this mumbo jumbo.

205
00:22:54,960 --> 00:23:03,950
But anyway, is it is just a couple of very quick questions for Russell.

206
00:23:03,950 --> 00:23:10,190
Yeah, you want to say, could I have one, too? Yes, I'll give you my card.

207
00:23:10,190 --> 00:23:16,220
How you got base much of this incredible process? Oh yes.

208
00:23:16,220 --> 00:23:25,250
Oh, you a question on. So I was sad when you were digging phase one and saw the amazing process by

209
00:23:25,250 --> 00:23:29,000
which you did it in two stages and stopped the surrounding buildings collapse.

210
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I just wondered if if the second phase was done exactly the same way, whether with a different process,

211
00:23:35,420 --> 00:23:40,070
it was done differently because somebody nearly died the first time round.

212
00:23:40,070 --> 00:23:49,640
It was quite so. They built the deck. They built the ground floor to prop open the hole because it's so deep, they actually collapses inwards.

213
00:23:49,640 --> 00:23:57,020
So. So one of the things is you actually prop the ground and form beams down the sides because it's such a deep basement.

214
00:23:57,020 --> 00:23:59,480
The second time around, they didn't do it that way.

215
00:23:59,480 --> 00:24:05,930
They actually did it in a more normal way, and they didn't go down sedate because you got that thing with people working underneath and on top.

216
00:24:05,930 --> 00:24:19,230
They were chasing the programme at the time, but is actually quite dangerous. Hello, John.

217
00:24:19,230 --> 00:24:24,390
I can appreciate what you did, I work with, they choke on an entire university.

218
00:24:24,390 --> 00:24:28,020
We were lucky because there was nobody there to object.

219
00:24:28,020 --> 00:24:35,400
But how do you get stuff into this building because I could barely get in on two legs if I was not take?

220
00:24:35,400 --> 00:24:39,320
How do I get in? It's a it's a question we ask in case Cambridge ever does this.

221
00:24:39,320 --> 00:24:47,800
Yeah. Oh good. Yeah, I'll give you my card. So a concrete purpose is the answer so that we could get.

222
00:24:47,800 --> 00:24:51,210
It was worked out quite carefully if you got smaller trucks.

223
00:24:51,210 --> 00:24:58,350
But then, for instance, the concrete they got in as far as they could from the gates at the top of the road, down the side here.

224
00:24:58,350 --> 00:25:04,620
And then they had rather great concrete. So a lot of it came in this new normal.

225
00:25:04,620 --> 00:25:13,020
Oh, right. So they come in. Yeah, so they come round the corner and the building is actually slightly counted for the turning of white vans.

226
00:25:13,020 --> 00:25:18,930
And then they're brought into the vehicle, into the storage. And all that comes into the side then either goes on out.

227
00:25:18,930 --> 00:25:21,810
But I think it's changed. I've actually come back and out this way.

228
00:25:21,810 --> 00:25:29,910
And so this bit of the corner, this bit, the building is actually cuts off at ground floor level for the ten circles in those vans.

229
00:25:29,910 --> 00:25:37,320
Rearrangement Yan will now Yan Lover will give her the next talk, but bacterial nanomachines?

230
00:25:37,320 --> 00:25:47,250
And then do we want Dale after you? Then you just sort of, oh yeah, Dale, do you want to be at the end or just off at the end?

231
00:25:47,250 --> 00:25:54,950
You want to be at the end, OK? OK, well, hopefully Christian will be ready after Jan and thanks.

232
00:25:54,950 --> 00:25:58,910
Thank you, Lydia, for organising this. It's wonderful.

233
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I really enjoy being here. It's sort of slowly sad.

234
00:26:02,750 --> 00:26:09,050
I know Dave will leave a huge hole in the field and sort of that's a good looking forward to that.

235
00:26:09,050 --> 00:26:13,590
But I can see so many talented people that have been trained really well.

236
00:26:13,590 --> 00:26:19,220
I will take over, and so it's really wonderful. So makes it a slightly more serious point.

237
00:26:19,220 --> 00:26:29,270
I don't think this has been mentioned so far. I think seeing all this amazing signs and all these people have become companies, that's quite unusual.

238
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The number of people, and I think it shows generosity on their part, sort of let people develop into their thing.

239
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And I think that's sort of a a real sign of a sort of great scientist,

240
00:26:41,660 --> 00:26:50,380
someone who sort of understands that, that it's about team effort and people being able to do their thing.

241
00:26:50,380 --> 00:26:55,680
As you probably know, most people have no I have not been in Dave's school,

242
00:26:55,680 --> 00:27:06,970
but we have worked together for many years and mostly on Facebook and Mark B.F. and and that's some of the stuff I will talk about today.

243
00:27:06,970 --> 00:27:16,570
This is old cell division, as you know, and sort of the I think the impact that we had has led to certain recognitions.

244
00:27:16,570 --> 00:27:24,130
One of them is that we were allowed to organise this workshop in 2009 here in Oxford, which which,

245
00:27:24,130 --> 00:27:31,270
you know, I enjoyed a lot initials of a number of people or so that became quite famous in that field.

246
00:27:31,270 --> 00:27:38,890
And I sort of I was really proud to be part of that.

247
00:27:38,890 --> 00:27:47,260
I wanted to give you a sort of one glimpse of a career defining moment for me that involved Dave and probably doesn't remember this.

248
00:27:47,260 --> 00:27:54,340
I was a young P.I. It was 2001, and we had just discovered that maybe is acting like.

249
00:27:54,340 --> 00:27:57,370
And this was sort of a great success,

250
00:27:57,370 --> 00:28:05,470
but it wasn't entirely sure whether I should stay with bacteria and prokaryotes because funding is not as easy and publishing definitely isn't.

251
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And so I went to an MBA meeting where Dave was invited as a sort of.

252
00:28:12,180 --> 00:28:20,250
Famous scientists to give advice to young people, and I woke up to them and sort of asking, what should I do and I should I stay with this country?

253
00:28:20,250 --> 00:28:27,880
He clearly advised me to stay with prokaryotes because it's the beauty of simplicity that often enables discoveries.

254
00:28:27,880 --> 00:28:33,590
And I have to say I do not regret following that advice, and it has been actually wonderful.

255
00:28:33,590 --> 00:28:43,850
So I'm going to talk about two things, one is after this, it's a story that was a recent one by Nicola Joy in my lap and this can be very brief.

256
00:28:43,850 --> 00:28:51,050
I just want to show you the beauty of this. This enzyme, it's part of the reason that's this complex that divides cells.

257
00:28:51,050 --> 00:28:57,710
And I want to sort of delve too much into that. Have to say has been introduced by effects already.

258
00:28:57,710 --> 00:29:04,220
It has a membrane that sits in the division within a so the plastic pot, that's a DNA pump.

259
00:29:04,220 --> 00:29:07,160
And again, this has been introduced.

260
00:29:07,160 --> 00:29:17,180
So if this k pommes DNA around until the decides line and a married chromosome, it's dissolved into through one of their chromosomes.

261
00:29:17,180 --> 00:29:22,940
It's a repair pathway, and it's it's really important.

262
00:29:22,940 --> 00:29:35,420
Now we worked on in the early 2000s and sort of in the middle 2000s and on the structural aspects of this and the mechanistic aspects.

263
00:29:35,420 --> 00:29:43,280
And this is sort of summarised here after these three domains, as I said, and the commitment to to assign alpha, beta and gamma domain.

264
00:29:43,280 --> 00:29:47,300
And this sort of a lot of details that I don't want to go into.

265
00:29:47,300 --> 00:29:51,920
One important detail is that the government domain is the part that actually gives the system

266
00:29:51,920 --> 00:29:57,800
directionality that these cops sequences that that tell the enzyme which vie on the DNA to go,

267
00:29:57,800 --> 00:30:05,870
whether left or right. And this is obviously very important. And that structure that we solved clarified this, I think, in great detail.

268
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What wasn't clarified at the time is how exactly the double stranded DNA translocation works.

269
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They aren't actually that many double stranded DNA trans cases and known most of them are packaging

270
00:30:17,690 --> 00:30:23,250
motors and viruses and phages and sort of in cells that are not actually needed that much.

271
00:30:23,250 --> 00:30:28,490
But this is one of them. That's a real difficult issue, and that's why this hadn't been done.

272
00:30:28,490 --> 00:30:33,300
It's bloody fast. It's actually one of the fastest ATP AIDS is known.

273
00:30:33,300 --> 00:30:38,940
It's definitely the fastest motor known. It has two and a half thousand per second.

274
00:30:38,940 --> 00:30:47,930
Now this is really fast and is an experimental section in an experiment where we looked at whether you can actually sort of look at the enzyme,

275
00:30:47,930 --> 00:30:57,940
but it works and look at the ATP tries. There's no ATP left in any of the of the time points if he cannot get this enzyme in

276
00:30:57,940 --> 00:31:02,640
a test tube while there's still ATP because it just hydrate absolutely everything.

277
00:31:02,640 --> 00:31:11,660
If you did discover by doing this experiment that it could become hydrolysed slowly and we can use that as a as an analogue to look at the enzyme,

278
00:31:11,660 --> 00:31:14,300
what it works, that's what we did.

279
00:31:14,300 --> 00:31:23,360
Khayyam come came to the rescue with so many things these days, and only the mice sample is asymmetrical and we presume is translocate.

280
00:31:23,360 --> 00:31:28,460
Everything else always has the same nucleotides in all six subunits, and it's not interesting.

281
00:31:28,460 --> 00:31:31,040
This just shows you the beauty of em.

282
00:31:31,040 --> 00:31:39,080
You can get very, very clean densities and objects like this, and this is the DNA in the middle here, and that's the f k domains left right.

283
00:31:39,080 --> 00:31:47,210
So this is showing the asymmetry. So you can clearly see that this is not a sort of a C6 circle and that, you know, it's it's a, you know,

284
00:31:47,210 --> 00:31:55,760
off the centre in the middle here is we guessed originally, many years ago and very importantly, the structure.

285
00:31:55,760 --> 00:32:03,050
The one structure shows an entire hydrolysis cycle you can see here that's the ATP to be hydrolysed ATP.

286
00:32:03,050 --> 00:32:06,320
Then this is in exchange for this sort of mix of HP and ATP.

287
00:32:06,320 --> 00:32:11,870
And this is the two entities and you can see that the hydrolysis here goes goes clockwise from the top.

288
00:32:11,870 --> 00:32:21,510
Very clear from just looking at one structure. And if you go on to the ring, the six minutes have had six different confirmations.

289
00:32:21,510 --> 00:32:26,140
Alpha subunit beta subunit here by a domain in beta domain,

290
00:32:26,140 --> 00:32:32,550
and you can see the six different states that are mostly described by the angle between these two domains.

291
00:32:32,550 --> 00:32:39,220
OK. And you will see in a minute why this is the key for the translocation of this enzyme.

292
00:32:39,220 --> 00:32:46,720
One more detail. So for subunits, units of four Peter domains are in contact with the DNA, that's asymmetric.

293
00:32:46,720 --> 00:32:55,230
Two of them are not in contact. And the whole thing is used to describe this spiral staircase to the settlements

294
00:32:55,230 --> 00:32:59,460
intact with the backbone of the DNA only because the sequence unspecific.

295
00:32:59,460 --> 00:33:06,240
And you can then basically follow the DNA with this molecule that's actually sort of symmetrical.

296
00:33:06,240 --> 00:33:10,110
So this is the movie, how all this works, that we first meet.

297
00:33:10,110 --> 00:33:14,160
It's so darn fifty thousand times so we can actually look at it.

298
00:33:14,160 --> 00:33:21,060
But each transportation step, the subunit each of the subunits is one of six confirmations and then be colour coded this year.

299
00:33:21,060 --> 00:33:27,040
So the domain that's blue is the one that extra hydrogen circles round and round and round and round in the circle.

300
00:33:27,040 --> 00:33:32,550
OK. You can also see that the DNA is is only gently rotating.

301
00:33:32,550 --> 00:33:42,030
This is a mismatch between the two base transportation step and the fact that DNA has 10 base pairs

302
00:33:42,030 --> 00:33:48,060
repeating something that so there's a slight rotation and the enzyme does not follow the key construct.

303
00:33:48,060 --> 00:33:53,430
So here I show you again, the six subunits in the different confirmations do this little dance.

304
00:33:53,430 --> 00:34:02,990
It actually describes a wave that is related to the felicity of the day because you need to need to go round as the backbone of that.

305
00:34:02,990 --> 00:34:06,710
The iPhone domains are more or less of a Pacific society device.

306
00:34:06,710 --> 00:34:12,620
They form a ring on the tops of the DNA probably never really sort of goes off centre.

307
00:34:12,620 --> 00:34:20,720
So then let's look at the DNA interactions in the ring. You can see it grabs the DNA there one two three four.

308
00:34:20,720 --> 00:34:26,330
Then it goes off and resets two steps down and then goes one two three four.

309
00:34:26,330 --> 00:34:33,470
That is why this for certain phones and the two subunits are the ones where you reset back so you can bind at the bottom.

310
00:34:33,470 --> 00:34:37,700
Super simple. Now this can actually be understood as Trutmann,

311
00:34:37,700 --> 00:34:45,830
so they can see that the red subunits basically form a filament along the DNA and then the other subunits respond to the other side.

312
00:34:45,830 --> 00:34:47,570
This is something we know from.

313
00:34:47,570 --> 00:34:59,320
So this little filaments and this sort of beautiful that it turns out that these transfer cases use something that we know from silicon systems.

314
00:34:59,320 --> 00:35:04,440
That's all I wanted to say about this game. So I move on to a the F.

315
00:35:04,440 --> 00:35:08,310
This is work done by Frank Berman is actually here, it's in the back.

316
00:35:08,310 --> 00:35:16,820
I also need to mention this on the job market, so if you want to hire him, please talk to him and to look at his publication.

317
00:35:16,820 --> 00:35:20,150
So the Colo chromosome is a giant molecule, it's, as you pointed out,

318
00:35:20,150 --> 00:35:27,650
already just to drive home that point that DNA needs to fit into that little point in order to be packaged.

319
00:35:27,650 --> 00:35:32,600
So this is a very strong connexion with actual compaction reaction that's needed and so be

320
00:35:32,600 --> 00:35:39,980
known for a long time that this involves somehow looping or a length versus condensation.

321
00:35:39,980 --> 00:35:49,970
S.A. closing complexes are ancient organisers of chromosomes, and they are involved in this and get sort of a very schematic view of how this works.

322
00:35:49,970 --> 00:35:52,970
They form this ring like structures. They're introduced already.

323
00:35:52,970 --> 00:35:58,460
It's a dimer or the hetero homo dimer and declines in protein that form a tripartite ring.

324
00:35:58,460 --> 00:36:02,060
And they go round to DNA and that can go around two days,

325
00:36:02,060 --> 00:36:09,230
either for cohesion that separates pieces of DNA or for condensation that the same piece of DNA.

326
00:36:09,230 --> 00:36:13,460
Now this is still a bit controversial. We don't entirely sure that this is actually what's happening,

327
00:36:13,460 --> 00:36:20,570
but I still like the simplicity of this scheme, and I still hope that that will turn out to be right.

328
00:36:20,570 --> 00:36:24,800
So this means that cohesion and condensation are possibly related.

329
00:36:24,800 --> 00:36:37,550
Activities that are caused by this topological entrapment of two days in a ring must be if it's the same complex of E. coli and it's a bit special.

330
00:36:37,550 --> 00:36:44,870
So here's a drawing of a typical SNC complex that has this long SNC proteins in the

331
00:36:44,870 --> 00:36:51,740
dimer and then the ATP's heads are linked with this crisis and here a cohesion and

332
00:36:51,740 --> 00:36:57,050
condense and have two subunits that bind to the cross and must be if one has won the

333
00:36:57,050 --> 00:37:05,780
Class A. F. And there's just this nomenclature in must be if it's a Homo dimers.

334
00:37:05,780 --> 00:37:14,590
But because the class act introduces a symmetry that can be distinguished, one is called me for neck and the other one is called cup of a cup.

335
00:37:14,590 --> 00:37:18,640
Dave has worked on this for many years and worked many details,

336
00:37:18,640 --> 00:37:23,020
and this is sort of a scheme that tries to summarise some of this might be

337
00:37:23,020 --> 00:37:28,570
if it's meant to load everywhere on the cell and condensed it for a session,

338
00:37:28,570 --> 00:37:34,450
but not in the tower region, where there are signs that bind too much, but then help unload it.

339
00:37:34,450 --> 00:37:42,000
And you would normally then think that a sort of nucleotide encoder looks a bit like that in the wild type situation.

340
00:37:42,000 --> 00:37:46,260
So question how does must be intact with DNA?

341
00:37:46,260 --> 00:37:54,300
How does it contents or loop DNA? And how does it unload? Just as a quick recap, we have a must be human DNA test decency.

342
00:37:54,300 --> 00:38:00,630
Protein is the closest. And this is dimer is well, don't remember this.

343
00:38:00,630 --> 00:38:06,070
You will get lost. This is just to show that we have a sample.

344
00:38:06,070 --> 00:38:11,940
So Frank put a lot of effort into this finding the right organism and doing this well.

345
00:38:11,940 --> 00:38:21,070
So just to mention this briefly. So this is a must be a vulnerability that could be two e for F, but it's actually it must be two missing.

346
00:38:21,070 --> 00:38:28,350
So thought. And then we had DNA with testing site on it and much dimer and some ATP.

347
00:38:28,350 --> 00:38:32,080
And then we do some outcomes and structure.

348
00:38:32,080 --> 00:38:37,710
It is. This is really complicated. So you can see a many things.

349
00:38:37,710 --> 00:38:43,470
The first thing to notice is that the heads are down here and the Cocozza go into school straight up next, default and elbow.

350
00:38:43,470 --> 00:38:48,000
This is something Frank discovered earlier, very neatly breaking symmetry.

351
00:38:48,000 --> 00:38:56,440
Don't get the homo domain. The other thing you immediately notice is this two DNA is this one down here and this one they're found too be.

352
00:38:56,440 --> 00:39:03,340
And then the Claassen has a complicated arrangement on here, so let me just try to start a movement to talk it through this quickly.

353
00:39:03,340 --> 00:39:10,340
So that's the couple and maybe that's the knee. That's the markets in orange that's mucky.

354
00:39:10,340 --> 00:39:16,820
It's a diner. Then there's nothing to talk about that has a beautiful pipeline block from that, there was my pee.

355
00:39:16,820 --> 00:39:21,860
This is the heads and this are two dinners. It's called the NEC.

356
00:39:21,860 --> 00:39:26,480
That's the larynx. This points to the DNA, and that's the joint that wants to the punch of DNA.

357
00:39:26,480 --> 00:39:30,410
That's the oval that folds. That's the hinge that demonises the to my face.

358
00:39:30,410 --> 00:39:36,320
And that is the arms. It's all the colour code. So that's that complex.

359
00:39:36,320 --> 00:39:38,360
As I said, there's two days there,

360
00:39:38,360 --> 00:39:46,590
and it's very important now to point out that these two journalists are fundamentally different in the way they are bound.

361
00:39:46,590 --> 00:39:53,160
So they the lower one is what we call the clamp, or it's clamped, the DNA is found on top of the head,

362
00:39:53,160 --> 00:40:00,630
as you can see there, the stop this here and you can see that the closer which an orange actually goes around the DNA,

363
00:40:00,630 --> 00:40:07,170
so the air is actually trapped here in this compartment where that DNA is trapped in the other compartment up there, right?

364
00:40:07,170 --> 00:40:14,190
So they are in two different circles that the protein forms. So there isn't just one circle just to show again how this is done.

365
00:40:14,190 --> 00:40:15,400
So the classic goes, Why don't you?

366
00:40:15,400 --> 00:40:22,770
You can actually, for the first time ever and second, this completely see that close, and there's no question where that goes.

367
00:40:22,770 --> 00:40:30,410
And we need to discuss these two components. Before do that, just a brief mention,

368
00:40:30,410 --> 00:40:38,870
because dinosaurs demise have been mentioned as some journalists work here just in dates that has been controversial for many years,

369
00:40:38,870 --> 00:40:43,490
but I think this is very clear now from his work and also from the structure that

370
00:40:43,490 --> 00:40:49,400
there is a dinosaur that is density here very clearly for a second Macduff.

371
00:40:49,400 --> 00:40:54,350
And this is how the shows going to show you moving to sort of go quickly through this.

372
00:40:54,350 --> 00:41:02,690
So you you take this one and must be a monument and then and then the dinosaur via the RFQ in the middle.

373
00:41:02,690 --> 00:41:07,490
And I think from this looking at how that's pretty quickly.

374
00:41:07,490 --> 00:41:14,060
So if you just look at that structure here, this is sort of almost a domain smoke, so the winged helix in the middle domain go across.

375
00:41:14,060 --> 00:41:20,960
This is not accidental. This is this is something designed like that. So Mark B.F. is clearly a diner or diner.

376
00:41:20,960 --> 00:41:26,570
So I told you there are two dinners and this is sort of look at that.

377
00:41:26,570 --> 00:41:29,570
What we can learn from that if you just look from the top here.

378
00:41:29,570 --> 00:41:35,750
So we're looking at the DNA that's gone to Marty and me looking at the DNA that binds to the heads and the clam.

379
00:41:35,750 --> 00:41:44,660
And it turns out that the angle between these two diners is exactly what you would get in a negative super called plectrum, and this is shown here.

380
00:41:44,660 --> 00:41:48,910
So. This could be coincidental. There's no question about that.

381
00:41:48,910 --> 00:41:56,530
But it could also be that it signals that since you would expect chromosomal DNA to be organised like that,

382
00:41:56,530 --> 00:42:04,840
that that's there's something much more this month, something deeper about this, this correlation than just looking at that and that way.

383
00:42:04,840 --> 00:42:09,220
What this means is because the DNA chromosome is circular.

384
00:42:09,220 --> 00:42:16,660
There's no open ends under normal circumstances. The structure we're seeing must look a little bit like this because there must be a loop.

385
00:42:16,660 --> 00:42:27,360
So we call this a double lock where the DNA is in the clamp down here and then loops back through the murky morass a constellation and goes out again.

386
00:42:27,360 --> 00:42:36,460
OK. Otherwise, you cannot get that into into that that molecule was sweating it in order to investigate whether this happens in cells.

387
00:42:36,460 --> 00:42:46,840
Frank adopted a very complicated and finicky essay that comes from Stefan Gruber's lab, which in itself is based on Kim's work on on cohesion,

388
00:42:46,840 --> 00:42:55,420
combined testing cross-linking with Celsius so that it can look for concentrated molecules.

389
00:42:55,420 --> 00:43:04,720
So the idea is that we introduce cysteine pairs into various places so that protein protein interactions become covalent sustains yet.

390
00:43:04,720 --> 00:43:07,780
This can be done in cells that becomes a disulphide.

391
00:43:07,780 --> 00:43:16,210
And then you can denature whatever you like and you end up with a cut, a covalent cultivation event that you can trust.

392
00:43:16,210 --> 00:43:21,760
OK, so and then you you do engineered this into cells.

393
00:43:21,760 --> 00:43:25,690
You do this all in cells allows the cells across blocks.

394
00:43:25,690 --> 00:43:35,290
You wash all of the protein that's not bound to the DNA. Degraded DNA and then recover the protein that was cut and dated to the chromosome,

395
00:43:35,290 --> 00:43:40,130
and it can integrate many things, but here are the ones that we did this the ring compartment.

396
00:43:40,130 --> 00:43:43,660
This is the yellow bit here crossing Croslin's hinge.

397
00:43:43,660 --> 00:43:51,280
Gosling's clam is the cross and cross links and the head crosslink and then the hinge and the head for the frame.

398
00:43:51,280 --> 00:43:56,790
For those who know this is Jake and Jess.

399
00:43:56,790 --> 00:44:03,380
They're not going to talk you through everything very quickly. So in the ring compartment, there is entrapment here.

400
00:44:03,380 --> 00:44:06,350
This is just a test, whether the proteins actually do what they're meant to do,

401
00:44:06,350 --> 00:44:16,140
but they don't get entrapment in a nucleotides enact a mutant that doesn't by nucleotide in the mutant, it doesn't hydrolysed nucleotide.

402
00:44:16,140 --> 00:44:20,100
The same for the clamp. You get entrapment in the clamp.

403
00:44:20,100 --> 00:44:27,120
This is a control here from before you get in trouble in the but you don't get it with all nucleotide binding or no protected policies.

404
00:44:27,120 --> 00:44:32,250
And now in the frame, you don't get entrapment.

405
00:44:32,250 --> 00:44:37,080
OK. Why is that the case, how does this make sense?

406
00:44:37,080 --> 00:44:40,700
And this is the slide that I need you to remember.

407
00:44:40,700 --> 00:44:48,340
So it's a trick, so let's just assume that everything looks like as we thought, it's it's this double lock arrangement.

408
00:44:48,340 --> 00:44:55,470
And we apply all the collated links that are just talking for the ring and frame, so here.

409
00:44:55,470 --> 00:45:02,850
One of the dangers still trapped in the ring compartment here, one of the DNA is still trapped in the clam compartment,

410
00:45:02,850 --> 00:45:08,250
but here in the frame, both of them can slide out because they're not in trouble.

411
00:45:08,250 --> 00:45:18,010
OK, so we take this as a demonstration that our structure reflects.

412
00:45:18,010 --> 00:45:26,440
In many ways, the arrangement that we see with the crosslinking in cells is interesting things you can do

413
00:45:26,440 --> 00:45:32,830
just to mention that the the pattern we see with the entrapment excludes things like cohesion,

414
00:45:32,830 --> 00:45:38,080
for example. So this cannot be to deny this must be done on one day, and I'm going to talk to that.

415
00:45:38,080 --> 00:45:42,950
So the double lock explains the in vivo entrapment is the conclusion.

416
00:45:42,950 --> 00:45:51,950
Interestingly, the same pattern has been reported for another bacterial SNC complex and also for cohesion.

417
00:45:51,950 --> 00:45:57,200
So, so they find exactly the same SKG and J.S. entrapment,

418
00:45:57,200 --> 00:46:03,920
which means that maybe all the SNC complexes have universal binding mode that looks a little bit like the double lock.

419
00:46:03,920 --> 00:46:08,630
This is supported a little bit by the fact that the structure was done of cohesion and condensed,

420
00:46:08,630 --> 00:46:13,430
and it marked the EFF in the clan state are actually very similar.

421
00:46:13,430 --> 00:46:18,320
Obviously, they don't have the second DNA bond, but that's not the point.

422
00:46:18,320 --> 00:46:23,720
So let me just say to the to the model, So how does it unload?

423
00:46:23,720 --> 00:46:31,040
We have these different structures in different states here. They're going all the way from a fully rebound to April that goes to the movie.

424
00:46:31,040 --> 00:46:40,430
You can see that the change here in the confirmation is such that both oxygenase one down here and one up here cannot bind.

425
00:46:40,430 --> 00:46:45,980
In the April form, which means that we think that sort of one of the states is the unloaded state,

426
00:46:45,980 --> 00:46:48,260
whereas the other one is in the process of unloading.

427
00:46:48,260 --> 00:46:59,660
And based on that, we can then make up a model that takes us into account where we think that you open the the gate down here,

428
00:46:59,660 --> 00:47:03,020
then the DNA can slip out that pulls down with it.

429
00:47:03,020 --> 00:47:10,430
They didn't show you that the whole thing comes out and then you just close the whole thing again in the April state and it has come out.

430
00:47:10,430 --> 00:47:18,180
So basically a model for how based on Inuktitut hydrolysis, you would get in this particular complex,

431
00:47:18,180 --> 00:47:29,440
multi directed unloading only when s on my mother's side on the chromosome, which explains why, only into so this was a bit fast.

432
00:47:29,440 --> 00:47:36,590
So. The double lock is interesting because it sorts the two strands of DNA parts of the complex knows which one is which.

433
00:47:36,590 --> 00:47:43,490
It's very important. It could be a productivity device, so the thing goes probably on four kilo basis of the kilo basis,

434
00:47:43,490 --> 00:47:49,490
and having this always locked in logically makes sure that this doesn't come off.

435
00:47:49,490 --> 00:47:54,320
And then it's not clear yet whether this is required for transportation exclusion policy.

436
00:47:54,320 --> 00:48:18,360
That's all I wanted to say. Thank you for running over. The questions they'll.

437
00:48:18,360 --> 00:48:24,090
So why is the whole thing a diamond then? Good question.

438
00:48:24,090 --> 00:48:28,590
There's another motive to them now. I think it's a very serious question.

439
00:48:28,590 --> 00:48:33,060
I think the the other question is why are the other SNC complexes, not timers?

440
00:48:33,060 --> 00:48:38,190
And how does this relate to their function? I don't have a really good answer to that. I think it is.

441
00:48:38,190 --> 00:48:47,910
It is intriguing. And I think it's important because some of the transportation models can be very easily explained in a dynamic context.

442
00:48:47,910 --> 00:48:56,020
And I think this time will help us to clarify that because there are certain constraints on directions and so on.

443
00:48:56,020 --> 00:49:01,290
So. But absolutely. Yeah, and that's a question from.

444
00:49:01,290 --> 00:49:09,750
So there's a question online. What is the evolutionary relationship between helicase and ask?

445
00:49:09,750 --> 00:49:13,970
How can this be applied to DNA sequencing? I don't know.

446
00:49:13,970 --> 00:49:19,700
I think you know what this is. No, no, sir.

447
00:49:19,700 --> 00:49:24,570
So that's very interesting. This is obviously a very deep relationship, but helicase is so the mechanism.

448
00:49:24,570 --> 00:49:29,010
The classification mechanism is related. This is actually related.

449
00:49:29,010 --> 00:49:34,260
All the triple A. like enzymes use something related to what I showed you.

450
00:49:34,260 --> 00:49:42,750
But but because double strand of DNA is a rigid substrate, it's it has is the cleanest implementation here because you can hang onto the substrate

451
00:49:42,750 --> 00:49:47,340
all the time and trust that your translocated at the next bit comes in the right place.

452
00:49:47,340 --> 00:49:58,540
Sequencing this is a really cool idea. It may be possible at some in some way to use this to do in cell sequencing would be This is fantasy.

453
00:49:58,540 --> 00:50:03,470
I'm. Just one last question.

454
00:50:03,470 --> 00:50:14,790
The idea that you have this loop inserted through the ring, the so-called double lock, and that that may be part of the loop extrusion process.

455
00:50:14,790 --> 00:50:22,130
I mean, how do you explain Case Decker's observation that the whole reason supposedly can and I just

456
00:50:22,130 --> 00:50:29,010
think maybe also condense and can loop extrude through DNA that has some quantum dot associated,

457
00:50:29,010 --> 00:50:38,750
well, some some large obstruction. But is depending dependent on which of the dangers is is translocated.

458
00:50:38,750 --> 00:50:47,000
So there's obviously the upper DNA in the complex, it's not unloading, so there's no much there would just be loosely associated,

459
00:50:47,000 --> 00:50:55,400
one would assume, whereas the lower one might, might be the one that sort of hangs onto the DNA in a more defined way.

460
00:50:55,400 --> 00:51:00,800
And so, you know, and that is where the dilemma, I think, could potentially come in that that you, you know,

461
00:51:00,800 --> 00:51:10,430
you could you could come up with a translocation mechanism that deals with that, but it does not explain this well for cohesion and contention.

462
00:51:10,430 --> 00:51:22,760
And you know, as you know and I know, published data on loop exclusion on cohesion says that you don't have topological entrapment or likely not,

463
00:51:22,760 --> 00:51:28,370
let's say, and that is contradicts what I showed you. But there's ways out of that.

464
00:51:28,370 --> 00:51:33,750
It's clear there's lots of stuff we can't explain. You may need new forces.

465
00:51:33,750 --> 00:51:38,840
OK, we should move on now to a question.

466
00:51:38,840 --> 00:51:44,810
OK. It's a I have to say it's a special treat for me to be here on this special occasion.

467
00:51:44,810 --> 00:51:48,710
So thanks Dave and Lydia and the organisers for having me.

468
00:51:48,710 --> 00:51:54,500
And I was really happy to be, and I should say too, to be given the opportunity to give this talk.

469
00:51:54,500 --> 00:51:59,540
Then came problems when I started considering what I should say today.

470
00:51:59,540 --> 00:52:06,070
So. I could I wasn't sure what should be the balance between talking about the science we are doing

471
00:52:06,070 --> 00:52:14,020
in my lab since I left the flap and I wasn't sure how much I should say about David Sharratt,

472
00:52:14,020 --> 00:52:23,410
the shallow columns of bacterial DNA dynamics. With this, as we said, outstanding research contribution.

473
00:52:23,410 --> 00:52:26,770
I wasn't sure I should talk about Dave,

474
00:52:26,770 --> 00:52:35,680
who likes the bare necessities of life and are sharing food and drinks with with friends and discussing about anything.

475
00:52:35,680 --> 00:52:45,310
But I thought, there's one thing maybe I can I can talk about, which is how to do a postdoc in Dave's lab in the 21st century.

476
00:52:45,310 --> 00:52:55,200
So as we said before that, Dave mentored maybe sixty six or sixty seven Ph.D. students and maybe twice as many postdocs.

477
00:52:55,200 --> 00:52:59,620
So we all have a different story to tell. But this is this is part of mine.

478
00:52:59,620 --> 00:53:08,740
So I said six years and they've got so I won't tell you about the old story and you know how it is with time only good memories remain.

479
00:53:08,740 --> 00:53:14,560
But then I need to tell you how it started. So I'm from what we call the French Connexion,

480
00:53:14,560 --> 00:53:22,750
which has nothing to do with the drug cartel and Dave's arrest as a drug dealer as a make revealed this morning.

481
00:53:22,750 --> 00:53:30,280
But it refers to the people that in France are working in microbiology and that are somehow connected to David Sharpe.

482
00:53:30,280 --> 00:53:37,900
So I did my Ph.D. with possible connexions in the room here. And ethics joined soon after that.

483
00:53:37,900 --> 00:53:48,160
And Mick was the director of the institute at this time. So Dave, purely for scientific reasons, would visit Toulouse quite often.

484
00:53:48,160 --> 00:53:52,830
And then in 2006, it was when I had finished my PhD.

485
00:53:52,830 --> 00:54:00,040
It was somehow a logic for me to to to go to him and to to have an interview for a postdoc position.

486
00:54:00,040 --> 00:54:04,660
So he said, OK, you want to join the lab? What's your project? I'm not sure that's perfect.

487
00:54:04,660 --> 00:54:14,260
You're welcome. And for some reason, for some of your reasons and against the advice of the entire French Connexion,

488
00:54:14,260 --> 00:54:18,330
I decided to go somewhere else to do my first postdoc in Paris.

489
00:54:18,330 --> 00:54:26,770
Three years after with no possible publication, she didn't go very well, I have to say I went back today when I wrote a letter today.

490
00:54:26,770 --> 00:54:32,260
Dear David John Sherratt, fellow of the I wasn't very proud of myself.

491
00:54:32,260 --> 00:54:37,210
Fellow of the Royal Society. I'm very interested about what you're doing in it.

492
00:54:37,210 --> 00:54:43,210
I have done this and I know they had a good laugh in the lab about this letter because he knew me, basically.

493
00:54:43,210 --> 00:54:48,790
But then she contacted me and said, Yeah, you were welcome three years ago and you're so welcome today.

494
00:54:48,790 --> 00:54:52,840
So that's the first thing that they did for me as a as a mentor,

495
00:54:52,840 --> 00:55:00,910
which is to give his trust and give me this second first chance to to join his love and to to work with him.

496
00:55:00,910 --> 00:55:07,600
And I think a lot of people in this room can say that Dave has changed our lives and and that for most of us,

497
00:55:07,600 --> 00:55:13,750
we don't know where we would be if it wasn't for this, these decisions he made.

498
00:55:13,750 --> 00:55:20,470
OK, so at that moment, I had a project which was to basically induce a great double strand breaks into

499
00:55:20,470 --> 00:55:24,880
the equal chromosome and see what happens in terms of chromosome dynamics.

500
00:55:24,880 --> 00:55:33,460
So in these days, we had a quite good vision of the choreography of the cell cycle during the chromosome, during the cell cycle.

501
00:55:33,460 --> 00:55:35,710
And then it was the time, as they would say,

502
00:55:35,710 --> 00:55:47,170
to modify the system and to see what's what's the what is the response of these, these these cell cycle processes?

503
00:55:47,170 --> 00:55:53,590
So I arrive with my partner on an issue, is touching a postdoc in Steve Bell's lab at the moment.

504
00:55:53,590 --> 00:56:01,630
Then we discover that Oxford can be a very dangerous place with all these nice pubs and nice people, and I arrive.

505
00:56:01,630 --> 00:56:08,260
I think that the rearrangement of the programme is quite nice because I arrived, they had moved in this the new biochemistry building already,

506
00:56:08,260 --> 00:56:15,970
and it was just at the moment of the 2009 annual workshop that Ian mentioned didn't stop.

507
00:56:15,970 --> 00:56:25,300
And then I started discovering that the people in the lab and Dave as a mentor, and there are two things here that I would like to mention.

508
00:56:25,300 --> 00:56:30,370
The first one is that I mean, it's been said in different ways, but that's the way I would put it.

509
00:56:30,370 --> 00:56:41,380
I think that Dave, I've always been and will always be a playful kid, so he likes to play with science to enjoy.

510
00:56:41,380 --> 00:56:45,640
He's very well disguised as an adult. We all agree on that point.

511
00:56:45,640 --> 00:56:51,850
But then, as Marshall said, we would spend some time playing with rope tubes and wires.

512
00:56:51,850 --> 00:56:57,700
Following these examples, we would see him coming down the stairs of the biochemistry building and asking people,

513
00:56:57,700 --> 00:57:03,400
You are maybe to hold the tube you are to pull like this.

514
00:57:03,400 --> 00:57:06,340
You are the reptilian that was always Rodrigo, the rescue zone.

515
00:57:06,340 --> 00:57:16,270
And sometimes we would we would manage to resolve a detonation links, for instance, with recombination events, and that was nice.

516
00:57:16,270 --> 00:57:20,230
Sometimes we would just end up entangled in DNA. But still, that was that was nice, too.

517
00:57:20,230 --> 00:57:25,780
Clearly, one lesson was that it doesn't hurt to have fun when you're doing science with.

518
00:57:25,780 --> 00:57:33,610
The second thing I want to mention, and I'm not sure it's appropriate yet, and I'm a bit surprised nobody mentioned it so well.

519
00:57:33,610 --> 00:57:41,650
That's that's to me the first time I would go to Dave's office and tell me about it, tell him about my results.

520
00:57:41,650 --> 00:57:49,270
Then it would sink in his chair and close his eyes. So I have to say that in the beginning, it's quite surprising.

521
00:57:49,270 --> 00:57:58,000
It's a bit unsettling. And yeah, we don't usually talk about that in public.

522
00:57:58,000 --> 00:58:02,320
But so in the lab, we used to call them nano nuts.

523
00:58:02,320 --> 00:58:05,380
It's like micro slits, but just shorter.

524
00:58:05,380 --> 00:58:15,190
And then after sometimes you just realise that it's some sort of meditation, she goes to another place to a higher state of consciousness.

525
00:58:15,190 --> 00:58:21,880
I'm sure that he can picture science like only some scientist can do.

526
00:58:21,880 --> 00:58:28,570
I think he's doing thought experiments. I'm sure he even does thought controlled experiments.

527
00:58:28,570 --> 00:58:38,050
And then you would open his mouth and always have the most brilliant and most relevant input, which is always surprising from someone sleeping.

528
00:58:38,050 --> 00:58:43,660
You would agree. And then that these are two things that I really enjoy every day.

529
00:58:43,660 --> 00:58:54,400
So I started working on the project. And so the basically the system was consisting of an ILC one side to induce double strand break,

530
00:58:54,400 --> 00:59:00,760
and this site was encompassed by DNA localisation system so that we could follow the localisation

531
00:59:00,760 --> 00:59:06,370
of both ends of the double strand breaks during the induction of the break and during the repair.

532
00:59:06,370 --> 00:59:14,020
And as Kim said earlier, the system is never perfect because you'd like to be able to induce the cut in only one system chromatin and not the other,

533
00:59:14,020 --> 00:59:17,470
and to to to try to do that.

534
00:59:17,470 --> 00:59:21,850
What we did was just having a less stable ISC,

535
00:59:21,850 --> 00:59:28,540
one enzyme with the dagger intact so that we could reduce the lifetime and find the induction conditions where

536
00:59:28,540 --> 00:59:37,690
you'd have the right amount in enough cells to to get this situation where only one chromatin is is broken here.

537
00:59:37,690 --> 00:59:45,370
So I would start playing with the system and I would go today after a few months and say, Well, then I think with this system, we can address this,

538
00:59:45,370 --> 00:59:52,390
we can address that, we can look at the resection, we can look at the dynamics, we can look at this with induction and all these brilliant details.

539
00:59:52,390 --> 00:59:56,860
And then, Dave, I think that's when you realise that I really need it to be channel.

540
00:59:56,860 --> 01:00:02,280
He looked at me and said. Christian, nobody cares about the details.

541
01:00:02,280 --> 01:00:06,120
I was shocked. Of course, we care about the details. I mean, we are scientists.

542
01:00:06,120 --> 01:00:12,390
But then I realised that what she was trying to tell me is when you start a project in science, then you really need to dream big.

543
01:00:12,390 --> 01:00:22,920
You really should aim at the biggest and most important biological question you think you can address with your new system and stick to it.

544
01:00:22,920 --> 01:00:32,700
And then that's what I did in my case, it was the search for the homology and the dynamic of Orakei, and after a while, as I mentioned earlier,

545
01:00:32,700 --> 01:00:41,980
we had a quite strong case to show that indeed, these 10 sisters could do the homology burying Nikolai than repair and segregate again.

546
01:00:41,980 --> 01:00:48,090
And this was all correlated to the formation with the formation of a huge structure of cricket.

547
01:00:48,090 --> 01:00:50,850
We called a recce bundle, which you can see here.

548
01:00:50,850 --> 01:00:59,730
So after maybe a couple of years, I went to his office with a draught short draught of the paper and then I would say,

549
01:00:59,730 --> 01:01:07,770
Look, they just did the big experiment. So what we think is, I think, is the most important burying a correlation with bundles.

550
01:01:07,770 --> 01:01:11,610
No details. He looked at me and said, You're wrong.

551
01:01:11,610 --> 01:01:24,540
The devil is in the details. So I think he was playing with my nerves and and actually at that time, we started rejecting a lot of data,

552
01:01:24,540 --> 01:01:29,880
which I had because I had done all most of the experiments I wanted to do anyway without telling him.

553
01:01:29,880 --> 01:01:34,080
And I had to go back to the bench to do additional experiments, of course.

554
01:01:34,080 --> 01:01:43,320
And most importantly, we spent hours in his office refining every word of the manuscript.

555
01:01:43,320 --> 01:01:51,560
And now that the pain has gone and I think I can say that it's the other way I've learnt the most with you,

556
01:01:51,560 --> 01:01:55,440
you could drive me crazy every day, several times a day.

557
01:01:55,440 --> 01:02:04,950
But as I said, the pain is gone, and now I realise that's what what basically you taught me is dream big and then do the hard work,

558
01:02:04,950 --> 01:02:14,100
do the hard work to get a piece of science which at least to your satisfaction is is worth it.

559
01:02:14,100 --> 01:02:19,140
OK. So following the details, of course, everybody cares about the details.

560
01:02:19,140 --> 01:02:23,340
Of course, the devil is in the details, and I think that following the details, that's what detectives do.

561
01:02:23,340 --> 01:02:25,620
And that's what they did during his career.

562
01:02:25,620 --> 01:02:34,110
Because I think been said by Gordon and Lawrence, who started working on horizontal gene transfer and the question of how this coded one plasmid,

563
01:02:34,110 --> 01:02:39,510
which cannot be transferred but on its own, can use another plasmid to be transferred.

564
01:02:39,510 --> 01:02:44,220
What are the factors that are required in system to transfer this mobilisation process?

565
01:02:44,220 --> 01:02:50,520
And maybe I'm wrong, but I'm thinking you're the one who proposed the term mobilisation or mobility of all.

566
01:02:50,520 --> 01:02:53,790
He doesn't even remember. Yes. Thank you.

567
01:02:53,790 --> 01:02:59,730
Thank you both. And then from this, you realise that this plasmid, they were forming dimers.

568
01:02:59,730 --> 01:03:07,710
The dimers needed to be resolved. Then this led him to work on recombination site site-specific recombination transposes

569
01:03:07,710 --> 01:03:14,010
and moving to chromosome dimer resolution and then segregation replication,

570
01:03:14,010 --> 01:03:16,740
as it's been said, repair and more recently,

571
01:03:16,740 --> 01:03:24,420
maintenance with the defences and following the details, she went from this to that with each time, I think,

572
01:03:24,420 --> 01:03:32,520
discovering more than what she was even looking for when he when, when, when he was following these.

573
01:03:32,520 --> 01:03:37,890
And when I was preparing this presentation, I realised that I did the exact opposite thing.

574
01:03:37,890 --> 01:03:43,560
I started from chromosome dynamics, then I moved to plasmid DNA conjugation.

575
01:03:43,560 --> 01:03:49,380
And I think it tells a little bit about the way topics come and go in science.

576
01:03:49,380 --> 01:03:54,960
And it's like, you're at your desk, you're reading articles, you pile them up on your on your desk.

577
01:03:54,960 --> 01:03:58,560
And after maybe every two or three decades or so,

578
01:03:58,560 --> 01:04:05,820
you can turn the file upside down and you start again and you realise that's important questions that were raised at the time.

579
01:04:05,820 --> 01:04:16,020
You've accumulated enough knowledge to actually address them. And that's what I think I tried to do when I started my lab in Leon six years ago,

580
01:04:16,020 --> 01:04:22,980
which was to have a fresh look at pathogenic conjugation to use what I had learnt during the study of chromosome dynamics,

581
01:04:22,980 --> 01:04:34,800
listen microscopy and apply this to the question of dynamic of DNA conjugation in life cells because we really don't know much about this.

582
01:04:34,800 --> 01:04:37,920
So just to remind you, conjugation is a DNA transfer process.

583
01:04:37,920 --> 01:04:46,380
It can transfer a plasmids or order the entire chromosome in some cases, and it's been discovered by Joshua and Tatsumi and 4:46.

584
01:04:46,380 --> 01:04:52,620
And then it was obviously a seminal nature paper. One page, no figures.

585
01:04:52,620 --> 01:04:56,640
Good old times, some would say and imagine, I mean,

586
01:04:56,640 --> 01:05:03,630
everyone immediately realised that it was a very important paper because it was the beginning of plasmid biology.

587
01:05:03,630 --> 01:05:05,670
They didn't know at that time that they were looking at plasmid,

588
01:05:05,670 --> 01:05:12,390
but then came a few a few years after and beginning of plasmid biology and, of course, bacterial genetics.

589
01:05:12,390 --> 01:05:16,180
Now we know a lot more about DNA conjugation.

590
01:05:16,180 --> 01:05:22,590
We know it's a contact dependent transfer that it's important for the evolution of bacterial genomes,

591
01:05:22,590 --> 01:05:29,900
and it's responsible for the dissemination of plenty of bacterial properties, such as drug resistance, for instance.

592
01:05:29,900 --> 01:05:34,130
And I'll skip this, but it's just to illustrate that when we started the project,

593
01:05:34,130 --> 01:05:41,750
we knew a lot about the molecular processes and the sequence of events that were going on in the cell,

594
01:05:41,750 --> 01:05:45,920
which is basically a recruitment of a complex with a very important protein.

595
01:05:45,920 --> 01:05:49,940
We call the relaxation that induce a here on the plasmid.

596
01:05:49,940 --> 01:05:55,400
And that's you the helicase activity to extrude extrude, sorry, the single strand of DNA.

597
01:05:55,400 --> 01:06:00,980
And then in the rest of it, you have secularisation of the single stranded DNA plasmid,

598
01:06:00,980 --> 01:06:10,730
then complementary strand synthesis and gene expression and conversion of this recipient set into what we call the transplant with new properties.

599
01:06:10,730 --> 01:06:21,110
And in the lab, we are interested in the role of the pilots in the dynamics of this transfer in the establishment and the expression of plasmid genes.

600
01:06:21,110 --> 01:06:29,330
And to do that, we first needed to develop a system to be able to look at the transfer of plasmids under the microscope in real time.

601
01:06:29,330 --> 01:06:42,530
So what we did that was to use the SFB, which is a single strand binding protein encoded by chromosome and labelled with a fluorescent protein.

602
01:06:42,530 --> 01:06:50,420
And this allows us to detect the single stranded DNA as it generates the donor in the donor and transferred into the recipient.

603
01:06:50,420 --> 01:06:56,900
And we can also have a report system to show us when has the recipient cell acquired the plasmid?

604
01:06:56,900 --> 01:07:04,220
And this is how it works. So we've got a plus meeting, which in our case is the plus name because it behaves very well and liquidate in the condition.

605
01:07:04,220 --> 01:07:12,810
We're using the lab and we put the parasites. The B binding protein, which will bind to this parasite, is only produced in the recipient.

606
01:07:12,810 --> 01:07:16,160
And, of course, the in absence of the parasites.

607
01:07:16,160 --> 01:07:23,480
But then when the cell receives the plasmid, all these diffusing bodies are recruited to the parasite.

608
01:07:23,480 --> 01:07:28,040
And then this tells us that the cell has received a double standard DNA.

609
01:07:28,040 --> 01:07:31,340
So this is how it looks in a microfluidic chambers.

610
01:07:31,340 --> 01:07:38,240
We've got the cells that are dark, that are done a certain point, and the plasmid and the recipients starts with diffuse fluorescence.

611
01:07:38,240 --> 01:07:47,780
And I hope you can see that as conjugation goes. You have all those cells that are that are actually receiving the plasmid.

612
01:07:47,780 --> 01:07:51,980
And from this basis, we can look at conjugation events.

613
01:07:51,980 --> 01:07:55,580
We can do single cell analysis of what happens in the donor.

614
01:07:55,580 --> 01:07:58,700
What happens in the recipient and look at I mean,

615
01:07:58,700 --> 01:08:07,850
we can basically ask a lot of questions by looking precisely at those three different population of donor recipients and transplant accounts.

616
01:08:07,850 --> 01:08:17,750
So in the beginning, with the STEM, we answer the very, very simple questions like is there a favoured entry point of the DNA in the recipient?

617
01:08:17,750 --> 01:08:24,560
And so we know it's impossible to be everywhere a little bit preferentially at the ball, but not so much.

618
01:08:24,560 --> 01:08:32,060
What is the rate of transfer? And we ended up with something which is around six or seven nucleotides per second,

619
01:08:32,060 --> 01:08:37,550
which is totally consistent with what have been determined by looking at the transfer of the entire chromosome.

620
01:08:37,550 --> 01:08:46,910
So sometimes you don't need fancy a microscope to do that. And we also observed that the conversion of the single strand of DNA clustered into

621
01:08:46,910 --> 01:08:51,680
double standard DNA is very fast in five minutes and the process is completed.

622
01:08:51,680 --> 01:08:58,850
So the first thing we did with the system was to look at the acquisition of resistance, and I go very quickly.

623
01:08:58,850 --> 01:09:02,660
But this work has been done by Sophia. She's in the room as well.

624
01:09:02,660 --> 01:09:09,020
I stole her from the lab. She was doing a product in the lab and then she came to my lab.

625
01:09:09,020 --> 01:09:13,320
So she is F1, F2 and F3 because she did.

626
01:09:13,320 --> 01:09:19,460
The Ph.D. with Francois was a huge success from the bust up with Dave.

627
01:09:19,460 --> 01:09:23,340
So that makes her F1 as well. And then a postdoc with me.

628
01:09:23,340 --> 01:09:30,320
So she's the three. I think she has the world record of of sherratt in Britain.

629
01:09:30,320 --> 01:09:38,570
OK, so again, we run this out and go very quickly. I'm sorry, but we ran this sort of experiments where you can see the acquisition of the plasmid,

630
01:09:38,570 --> 01:09:44,930
which is now green and in the cells that have acquired this tetracycline resistance encoding plasmid.

631
01:09:44,930 --> 01:09:56,090
You can quantify the production of the resistance vector, which is the tenth a point that is needed to confer resistance that can influence them.

632
01:09:56,090 --> 01:10:03,230
That will basically pump out the antibiotic from the cell, and you can quantify that in real time.

633
01:10:03,230 --> 01:10:06,710
And this is the kind of things we have for single cells.

634
01:10:06,710 --> 01:10:15,110
And if you consider the 20 minute maturation time of the funeral, then you realise that basically the gene is expressed in the protein,

635
01:10:15,110 --> 01:10:19,930
the resistance to produce as soon as the plasmid is available in that those kind DNA.

636
01:10:19,930 --> 01:10:29,540
You can do that as well in snapshots in time, so you mix the cells and every hour you'd look at at the population.

637
01:10:29,540 --> 01:10:33,530
The the the the the the eve extend inside the fence convenience,

638
01:10:33,530 --> 01:10:38,990
and then I think it became interesting when we were when we did the same experiment in prison,

639
01:10:38,990 --> 01:10:43,610
sophisticated inhibitor of protein synthesis and to our surprise,

640
01:10:43,610 --> 01:10:48,860
we observed that those cells are perfectly able to acquire tetracycline resistance anyway.

641
01:10:48,860 --> 01:10:55,700
So the question became how can cells in which protein synthesis is inhibited?

642
01:10:55,700 --> 01:11:02,240
How can they still produce the influx? Then, after acquisition of the plasmid, there was some sort of conundrum there,

643
01:11:02,240 --> 01:11:06,110
and then we realised that it's actually the case with many different antibiotics.

644
01:11:06,110 --> 01:11:15,500
You can treat the cells during conjugation with antibiotics that inhibit reputation of transcription or protein synthesis,

645
01:11:15,500 --> 01:11:21,740
and healthy wild type recipient would be able to acquire the plasmid and develop the resistance.

646
01:11:21,740 --> 01:11:30,920
So we turned to the A.C.O.R.N. system, which was actually known to be able to have low activity of E-flat,

647
01:11:30,920 --> 01:11:35,360
of a range of of different toxic compounds and antibiotics.

648
01:11:35,360 --> 01:11:41,570
And we showed that to do this, the cells need to the activity of this complex.

649
01:11:41,570 --> 01:11:50,780
So basically, what we showed was that ACR 30 does need flex activity, which is not sufficient for the cells to survive.

650
01:11:50,780 --> 01:11:55,510
But if they acquire the plasmid from a donor, then.

651
01:11:55,510 --> 01:12:04,210
There is a low level of activity of these fundamental processes, which is still enough to develop the resistance.

652
01:12:04,210 --> 01:12:11,800
So with this, I'll stop here. I will just show the people from the lab.

653
01:12:11,800 --> 01:12:22,210
So they meet your grand grandchildren in science so you can see that I'm doing my best for women in science.

654
01:12:22,210 --> 01:12:26,500
And here are the full Moon members, the collaborators here and the funding.

655
01:12:26,500 --> 01:12:29,650
And the last thing I wanted to say is, Dave,

656
01:12:29,650 --> 01:12:38,840
if you still have some interesting DNA conjugation or just in wine tasting in Leo, for instance, you're always welcome.

657
01:12:38,840 --> 01:12:43,150
We have a bench ready for you, Olivia, as well.

658
01:12:43,150 --> 01:12:51,310
And I would be happy to take you around and and try to somehow return what you've given me and maybe ask as well.

659
01:12:51,310 --> 01:13:01,540
So thank you for your attention. Christian, thank you very much.

660
01:13:01,540 --> 01:13:06,610
That's lovely talk and that we we it all worked out.

661
01:13:06,610 --> 01:13:19,730
Any questions? That's fine, I think that the an hour.

662
01:13:19,730 --> 01:13:26,420
OK, well, in which case, well, there is this one.

663
01:13:26,420 --> 01:13:34,720
Yeah, there. It's nothing.

664
01:13:34,720 --> 01:13:41,090
I think one thing that microbes do is they always encounter very low levels of antibiotics now in environment.

665
01:13:41,090 --> 01:13:49,900
So you say they see this background effect of these plants? And I mentioned earlier on about the idea of heavy metals, for example, about.

666
01:13:49,900 --> 01:13:56,540
Do you think this is a general system whereby if the organism you know, you get the mobilisation barely on it,

667
01:13:56,540 --> 01:14:01,760
then you've got this period of sustainability before they're actually intoxicated.

668
01:14:01,760 --> 01:14:03,680
So that's a very interesting point.

669
01:14:03,680 --> 01:14:11,390
What we know is that at the concentration high concentration of antibiotics, this activity is clearly not enough for the scientists, right?

670
01:14:11,390 --> 01:14:16,880
But that indeed, in rivers, for example, an antibiotic contaminated rivers or soil,

671
01:14:16,880 --> 01:14:23,900
we could imagine that this low-level activity is enough for somehow contribute to

672
01:14:23,900 --> 01:14:31,370
the dissemination of of specific resistances that are carried by genetic elements.

673
01:14:31,370 --> 01:14:43,460
And we've shown also in this work that when natural isolates or clinical isolates, you find new mutants of repressors of the HRA be toxic.

674
01:14:43,460 --> 01:14:47,630
And so you they just have an overproduction of CRT.

675
01:14:47,630 --> 01:14:55,640
And we showed that these cells are indeed even more efficient at acquiring the resistance from the from the donor.

676
01:14:55,640 --> 01:15:02,390
Christine, thank you very much. Thank you. We've got to move on to the last thought.

677
01:15:02,390 --> 01:15:09,210
Well, we know with new machines that mess with nucleosomes.

678
01:15:09,210 --> 01:15:15,090
OK, well, it was a delight to be asked to come to give a talk at this event.

679
01:15:15,090 --> 01:15:22,200
I prefer to think of this as an event to to we have something different going on.

680
01:15:22,200 --> 01:15:25,990
I prefer to think of this as an event to mark David's achievements rather than his retirement.

681
01:15:25,990 --> 01:15:35,300
I'm sure, as we all suspect, that retirement will be a generous description of what will happen over the forthcoming months anyway.

682
01:15:35,300 --> 01:15:47,070
Oops, sorry. The button. OK, so we've just got this right.

683
01:15:47,070 --> 01:15:49,170
So when you asked me to give us a title,

684
01:15:49,170 --> 01:15:55,920
I said I thought about what I was going to do when we'd all had full stop so many times that we were coming up with things,

685
01:15:55,920 --> 01:15:58,500
looking at different times what I was going to talk about.

686
01:15:58,500 --> 01:16:03,840
So I planned originally to talk about this, which is work that's going on in my lab, which, as the name suggests,

687
01:16:03,840 --> 01:16:08,280
these are large chromatin remodelling complexes that are involved in shifting nucleosomes

688
01:16:08,280 --> 01:16:13,320
and then changing your concerns and doing various things all relating to DNA repair,

689
01:16:13,320 --> 01:16:19,140
which is what my lab is interested in. Of course, Dave doesn't give a damn about any of these things if bacteria don't have his time.

690
01:16:19,140 --> 01:16:22,830
So. So I thought, well, maybe I should think about something else.

691
01:16:22,830 --> 01:16:29,910
I also then thought, Well, let's have a look and see what there is obviously going to say a bit about my interactions with Dave over the years.

692
01:16:29,910 --> 01:16:38,130
So many people, we've obviously seen this picture. Many people have read this article, I'm sure, which describes Dave's career history.

693
01:16:38,130 --> 01:16:45,370
Now the trouble with that is that it's written by Dave. So, of course, it's somewhat a glorified view of what actually happened.

694
01:16:45,370 --> 01:16:54,480
Of course, that's one. That's one version of events, but I'm going to give you another one because I had to look to see what was on the internet.

695
01:16:54,480 --> 01:16:57,330
And I found something else.

696
01:16:57,330 --> 01:17:09,810
So it turns out that Disney have a franchise that they've been developing for a while, and that it turns out that Dave's career is the latest version.

697
01:17:09,810 --> 01:17:20,650
So this will just run through for a moment. So they're going to document they're going to document its entire career history.

698
01:17:20,650 --> 01:17:33,620
So it's important you follow what's going on here because this is describing, first of all, Dave.

699
01:17:33,620 --> 01:17:41,650
And then they see there's a thread running through which concerns somebody else.

700
01:17:41,650 --> 01:17:46,090
OK, so I thought was great, but of course, everything's been hit by the COVID crisis.

701
01:17:46,090 --> 01:17:49,510
And so when I contacted Disney to find out how things were going,

702
01:17:49,510 --> 01:17:53,440
it turns out we're working from home at the moment, so progressive, but it somewhat delayed.

703
01:17:53,440 --> 01:17:58,150
So we have at the moment is is the opening credits for what's going to be in the movie.

704
01:17:58,150 --> 01:18:01,660
So but well, they did do is let me have a couple of things to help.

705
01:18:01,660 --> 01:18:06,220
They said we'd be happy to publicise this new movie that's coming. So they gave me two things to help.

706
01:18:06,220 --> 01:18:13,040
One was getting a storyboard, which I can now use so the the quality the graphics will be rather worse than it was.

707
01:18:13,040 --> 01:18:32,260
And I also. I lost my microphone here.

708
01:18:32,260 --> 01:18:34,120
OK, so.

709
01:18:34,120 --> 01:18:42,790
This is where the story begins, and this is a photograph I obtained from a source I won't name, but we can see somebody sitting here in the front row.

710
01:18:42,790 --> 01:18:46,390
I hope it was him. I was trying to work out which one it was going to be.

711
01:18:46,390 --> 01:18:53,330
Somebody else now. OK, well, you could tell me which one it is afterwards.

712
01:18:53,330 --> 01:19:01,870
Lydia thought it was you. Anyway, so as we heard, he's keen on trying to find out what's going on.

713
01:19:01,870 --> 01:19:08,440
So he set out to travel the universe to answer this quest to find out about the answer about his travels first took him to Manchester.

714
01:19:08,440 --> 01:19:16,940
As we know, it's a rather wet place. We all know about whether that anybody's ever been there.

715
01:19:16,940 --> 01:19:25,990
And they learnt many things there, developed skills that would serve families troubles can tell from this document that I.

716
01:19:25,990 --> 01:19:30,990
So. So the first place he went off to Manchester was up to Edinburgh.

717
01:19:30,990 --> 01:19:39,330
And this was even further north, but at least it rained less there. So and here is interested in interacting in many aspects of the force.

718
01:19:39,330 --> 01:19:42,570
He was strong, going strong and his Jedi skills are still in their infancy.

719
01:19:42,570 --> 01:19:47,820
So you need to go somewhere else to build on his strength and understand more about that.

720
01:19:47,820 --> 01:19:54,000
Meanwhile, at the same time, that little boy that we knew we've heard about earlier was an infant that was just starting school,

721
01:19:54,000 --> 01:19:58,060
so he was beginning his own journey of discovery.

722
01:19:58,060 --> 01:20:07,020
So she decided to encounter a new religion with an element called Collie, one that held secrets of life force and set off on a planet.

723
01:20:07,020 --> 01:20:12,430
The stuff that separate stuff since us and where we call California the natives here is strange.

724
01:20:12,430 --> 01:20:16,540
They listen to something called hippie music and worship the Temple Woodstock,

725
01:20:16,540 --> 01:20:23,800
where they took a mind expanding medications and intoxicating potions they believe brought them closer to the life force.

726
01:20:23,800 --> 01:20:31,170
So I wanted to blend in. He grows hair long, followed their ways and then became like them.

727
01:20:31,170 --> 01:20:41,550
This land is also inhabited by wise prophets that they've came across and some less wise prophets,

728
01:20:41,550 --> 01:20:48,870
but we've learnt much from these people and over the years a lot to think out of the box with very characteristic about the way they things.

729
01:20:48,870 --> 01:20:57,270
People hinted at this so far. It's a very long, short recollection, always crop ideas and a very good, long wonder where these good ideas.

730
01:20:57,270 --> 01:21:01,770
So anyway, after just two years, he's called back to the Planet Earth to bring its new coli,

731
01:21:01,770 --> 01:21:06,070
one beliefs with him and hoping to continue to worship its properties.

732
01:21:06,070 --> 01:21:09,910
So it's back to Earth, and he was summoned to the Sussex Academy on Planet Brighton,

733
01:21:09,910 --> 01:21:14,870
which is the strange world of sand and struck curious buildings while continuing

734
01:21:14,870 --> 01:21:20,140
worship the one place where the woman is and its ability to replicate itself.

735
01:21:20,140 --> 01:21:29,750
He also started to uncover a darker aspect. Stories emerged about a newly discovered source of DNA that was mystical.

736
01:21:29,750 --> 01:21:31,110
This is called a transposon.

737
01:21:31,110 --> 01:21:38,960
It was able to jump into other DNA elements, replicate itself and then jump out, but in the process that copied itself into something else.

738
01:21:38,960 --> 01:21:45,780
Actually, one just replicated itself in its own image. But the transposons are replicate themselves inside something else.

739
01:21:45,780 --> 01:21:50,430
So this is starting to get a little dark. So the dark forces dark side, the force is coming.

740
01:21:50,430 --> 01:21:55,020
Meanwhile, that little boys now still going home to school,

741
01:21:55,020 --> 01:22:01,950
oblivious of what was going to be happening to them in the future, and he was happy fishing and enjoying his life generally.

742
01:22:01,950 --> 01:22:10,140
Next, we find that the sheriff was recruited to a distant planet, Glasgow, in the nebulous Scotland Yard to develop his studies.

743
01:22:10,140 --> 01:22:21,570
This was a strange place full of snow capped peaks with a brief season that was known locally as summer and is training.

744
01:22:21,570 --> 01:22:29,910
Manchester obviously helped him very well and getting ready for this process. But it's also new challenges that came with moving to this new place.

745
01:22:29,910 --> 01:22:38,970
The native spoke with a strange tongue. You have to say it phonetically to understand.

746
01:22:38,970 --> 01:22:45,390
They also spoke to each other and can communicate with strange symbols.

747
01:22:45,390 --> 01:22:49,690
But he learnt their language and the secrets of their night to develop this new religion or new aspect,

748
01:22:49,690 --> 01:22:55,780
and they called site-specific recombination of the show. It was very keen to pass the word about their new newfound discovery.

749
01:22:55,780 --> 01:23:02,610
So you travelled again back to the North American continent to do this, and such as culturally.

750
01:23:02,610 --> 01:23:09,860
However, in this particular year here. And you can see him using his intoxicating potions to try and corrupt one of the locals,

751
01:23:09,860 --> 01:23:12,680
so these ideas of his new site-specific recombination religion,

752
01:23:12,680 --> 01:23:22,110
which some going to heaven but it can recognise who the lady is that I can figure with.

753
01:23:22,110 --> 01:23:28,720
And many found this new religion too hard to understand and resisted turning to the dark side.

754
01:23:28,720 --> 01:23:36,010
So then that was actually a day for worshipping Colby, one also uncovered something else called VR system.

755
01:23:36,010 --> 01:23:45,130
This was able to be understood using the symbols he learnt from his time in Glasgow and what you've done from transposons.

756
01:23:45,130 --> 01:23:48,910
But what did the system look like and what was this strange system?

757
01:23:48,910 --> 01:23:56,600
And he really wanted to see more of it to be able to visualise this. Meanwhile, in another place again and again in York,

758
01:23:56,600 --> 01:24:03,540
a young man was learning the skills of the night and how to use X-rays to study the secrets of DNA.

759
01:24:03,540 --> 01:24:10,230
So finally, by this point, the sheriff's abilities have been recognised by the other academy that are older,

760
01:24:10,230 --> 01:24:15,890
and he was recognised as a full Jedi master in 1992.

761
01:24:15,890 --> 01:24:20,930
And which led to its recruitment to the Oxford Academy and later in a few, just a few years later.

762
01:24:20,930 --> 01:24:27,590
This was a different world again. We're here. The peaks are not covered in snow, but we're made of ivory.

763
01:24:27,590 --> 01:24:35,030
He set up his church recruiting disciples to learn in many ways of the day religion that we've already heard how many strains.

764
01:24:35,030 --> 01:24:39,500
But he's still young to more about. Learn more about the axial proteins. So here we go.

765
01:24:39,500 --> 01:24:45,830
Meanwhile, a new recruit that arrived at the Oxford Academy only the year before, so when the show arrived,

766
01:24:45,830 --> 01:24:53,010
this post was already a follower of the faith and believed he could use X-rays to understand the molecular properties.

767
01:24:53,010 --> 01:24:56,230
The sheriff sought to corrupt him into the ways of site-specific recombination,

768
01:24:56,230 --> 01:25:04,040
an introduction to the zero proteins assuring this would be a dead shall easy project to work with the crystal structure.

769
01:25:04,040 --> 01:25:10,690
Many years later, they finally managed to solve the structure, and then that was revealed to them.

770
01:25:10,690 --> 01:25:15,530
But the ex-Chelsea protein of this day still remains hidden from view.

771
01:25:15,530 --> 01:25:19,700
But they managed to publish their work, so they were happy about that, or at least the young man was,

772
01:25:19,700 --> 01:25:26,690
he was very happy to be publishing this, especially with Dave, until he learnt that David already got 200 publications.

773
01:25:26,690 --> 01:25:31,700
So this was really rather insignificant contribution to the overall process,

774
01:25:31,700 --> 01:25:36,950
somewhat dispirited and think they would never be able to escape if he was forced to find the Excel surprising,

775
01:25:36,950 --> 01:25:46,940
given how long it took to be, the man left to join the rebel band of Jeddah is itself a little known remote location on the M25.

776
01:25:46,940 --> 01:25:56,500
However, he'd learnt much from the sheriff's teachings. And for many years afterwards, a young man to be can continue to be inspired by his wisdom.

777
01:25:56,500 --> 01:26:09,700
Meanwhile, continued, but came away from the camera away from the dark side and started to move into.

778
01:26:09,700 --> 01:26:15,580
Starting to move into other other areas and looking at wholesale instead of what we've been doing for the course,

779
01:26:15,580 --> 01:26:21,220
you can see he's using many lightsabers to actually detect the molecules within those cells.

780
01:26:21,220 --> 01:26:31,300
And as his powers grew stronger and stronger, the Wellcome Trust kept funding well beyond any sensible working age.

781
01:26:31,300 --> 01:26:38,620
Finally, various powers have become so great the Oxford Academy threw him out and the had to himself to the Remote Asteroid Chamber,

782
01:26:38,620 --> 01:26:44,110
where he now roasts local fauna and continues to drink intoxicating fluids.

783
01:26:44,110 --> 01:26:48,520
But we all know his story is far from over, but mine is, so that's the end of that.

784
01:26:48,520 --> 01:26:52,750
I just need to acknowledge various people who helped me get this together.

785
01:26:52,750 --> 01:26:56,900
And then, of course, finally. Thank, Dave. So thanks, Dave.

786
01:26:56,900 --> 01:27:05,090
But of course, now we know what his true identity is. We can all go and have a drink.

787
01:27:05,090 --> 01:27:10,430
No questions. We got to break. Dale, thank you very much.

788
01:27:10,430 --> 01:27:16,790
That's wonderful finale. And I'd just like to take this time to just thank all of you for coming.

789
01:27:16,790 --> 01:27:26,960
It's been a wonderful day as a great climax, and they've, you know, just keep, you know, being here and here tomorrow, I'm sure.

790
01:27:26,960 --> 01:27:35,060
So, so just remember there's a reception here, but we need to be in Trinity by seven o'clock.

791
01:27:35,060 --> 01:27:40,970
I think everyone knows where to go. So, yeah, thank you all very, very much.

792
01:27:40,970 --> 01:27:47,304
Wonderful day.