Royle, Stephen: Kate's just reached 12:00 o'clock, so it's time to start today's noble lecture. So I'm going. Perkins. I'm the Dean of medicine here at Warwick Medical School. It's my absolute delight that we've been joined by Professor Stephen Ward, who will deliver his inaugural lecture as a professor being with professor at the University of Warwick and particularly at Warwick Medical School is a really prestigious accolade. It really sets people in their academic career at the top of the the pinnacle at the top of the game.
Royle, Stephen: The concept behind it is an awful lecture is for the individual to share their work across their academic career with the wider community so that we can appreciate some of the absolutely fantastic things that are taking place here at Warwick Medical School.
Royle, Stephen: Stevens, a cell biologist in the Centre for Mechanical Chemical cell Biology and Andrew McCain, is still trying to explain to me what a mechano chemical sound biologist does, but I'm hoping that after Stevens lecture.
Royle, Stephen: I'll. I'll have you know greater clarity in with, you know, what a fantastic site title of a very cellular song is going to bring together. The division of Amoebi or small small creatures.
Royle, Stephen: With with singing so you know, 22 passions together. Stephen, very much looking forward to your lecture. And on behalf of the school, many congratulations in your chair.
Royle, Stephen: ****, I didn't ask about the lights. How do I dim them?
Royle, Stephen: I'm not.
Royle, Stephen: Do that. OK, alright. We'll just go. So I thought I'd start by explaining what the title of my lecture today is all about. A very cellular song. So it's actually a track of incredible string bands 1968 album, The Hangman's Beautiful Daughter. That's the front cover there. And it was suggested to me actually by my friend Sally Lowell at University of Edinburgh. So the song. Don't worry, I'm not going to play. It's 13 minutes long. So it would take too much time, but it's about how an amoeba divides. So I'm going to tell you today about cell division.
Royle, Stephen: Along with other things that sales do and give you a bit of biographical background. So that's the other reason that I picked it. One of my other passions besides cell biology is music. And so the two things coming together are quite nice. I actually the understanding cells and cell biology and what cells do is a little bit like understanding the cell's own songs. So I'll tell you today about different processes that cells do. And you'll see that they are the sum of lots of different proteins and molecules coming together as an ensemble.
Royle, Stephen: To orchestrate these functions, which are very much like cellular songs, so we're going to hear some of these today.
Royle, Stephen: I'd like to begin by thanking the folks in my lab over the years. This is some, but not all of them. They're happy smiling for the camera. Without them, I wouldn't have anything to present. They're the ones doing the hard work, generating the data that I'm going to show. I don't have time to go through everyone's work, but I just want to thank them for for their hard work. Actually, as a, as a π.
Royle, Stephen: You each year you get older and more wiser's, but these people it's a bit like Dorian Grey. These people stay forever young. They're always at the same stage of their careers coming into the lab, not knowing very much and hopefully leaving, knowing a bit more. And really that's one of the joys of the job is to see these people grow and develop as scientists. And that's a, you know, the cellular songs are one reason for getting out of bed in the morning. But my trainees are the other reason. So thank you to them and I'll. I'll thank them again at the end. So let's just have a little peek at some of the things that cells do.
Royle, Stephen: These are full movies taken by folks in my lab here in Warwick and on the left you'll see cell division happening and on the right hand side you'll see some membrane traffic and we're going to hear about both of these today. So on the left, we've got some. These are all human cells, a few down microscope on the left, you'll see a cell dividing. So it's sharing the the copied genome to the two daughter cells through the process of mitosis. And that's the mitotic spindle that you can see segregating the chromosomes. So the two daughter cells.
Royle, Stephen: On the right you can see membrane traffic. This is the process of putting proteins into little vesicles and sending them to different places in the cells that they can do other jobs. And really I work on this really for the intellectual curiosity of trying to figure out how these processes work. And I'll tell you about some of the nuts and bolts today. So that's the main goal. But the other thing to say is that there's a real impact here because all diseases have a cellular origin.
Royle, Stephen: And defects in these processes lead to human disease. Actually, if there's if there's a problem in either of these things in cells, then it's game over for the cell first and foremost. But if we just consider cell division on the left, the uncontrolled cell division is the cause of cancer. And so we're interested in how cancer develops. And also if we can target cell division as a way of inhibiting the growth of a tumour. So that's one kind of health impact. And on the right.
Royle, Stephen: Protein's been in the right place at the right time, underlies lots of other cellular processes, and defects in membrane traffic underlie lots of diseases such as nerve degeneration or blood clotting. There's there's a big long list and so understanding how these things work and how they go wrong in disease is a big motivation. OK, so, so that's the beauty of cell biology in one slide and something less beautiful. Now, I'm going to just.
Royle, Stephen: Begin at the beginning, which is.
Royle, Stephen: A me.
Royle, Stephen: And so, yes, So what? You probably can't tell here. So I didn't really have a passion for science right at this point. My passion, really, maybe you can see it. My eyes was heavy. Psychedelic music.
Royle, Stephen: So I I really pursued music as a as a my main interest really. I few years after this I formed a band and we, you know, I was trying to to do music and and actually.
Royle, Stephen: I was saved from a career in music by a not being good enough and B actually the the music industry collapsed at the end of the 1990s, so it's perhaps a good that I changed direction, but actually ultimately led to my career in science. So I actually got onto my band, split up by concentrated on my studies and got into University of Sheffield to do biology and.
Royle, Stephen: One day I was in the lecture and the lecturer said if you're interested in doing some research in the drug industry, come to this room at this time and I didn't actually have an idea what I wanted to do after I left university. So I went along to this room and the the guy was actually unusual. He'd worked in the drug industry and then come back to academia, and he was passionate about students getting some research experience. So he handed out some application forms, and I dutifully mailed them off and got several rejections. But I got one offer of a of an interview here at Black, so welcoming Stevenage.
Royle, Stephen: And actually the guy that interviewed me told me it's in my face and you know, you undergraduates. You're all the same. You've all got good grades and you're all a good universities. And it's very hard to pick you apart. But you've written on your CV that you're into music. And my wife plays the cello. So I thought I'd ask you for interview. And literally that was the. That was my big break.
Royle, Stephen: So I got the job and and I spent a happy year there. I I learnt a lot here. I learned that I was really, you know, research was what I wanted to do for the rest of my life.
Royle, Stephen: I also learned I didn't want to do that in the drug industry, so it really was informative. It was also the place I published my first first author paper, and it really, you know, I sort of went back to university with a renewed passion to finish off the degree and go and do a PhD and and and, you know, sort of do research, which I'd learned was so much fun. And I couldn't. I couldn't believe it. They actually paid me to to do this. It was so good. So I returned, finished my degree, and then got a pH. D place at Cambridge.
Royle, Stephen: In the lab of Ruth Mel Agnardo, who's the woman on the left.
Royle, Stephen: She was running a neuroscience Physiology lab.
Royle, Stephen: So I joined and and didn't really do any electrophysiology. I ended up doing some cell biology, so I'll just briefly explain this because it sort of sets the scene for what I did afterwards.
Royle, Stephen: So yeah, so this picture on the bottom left is me in the lab actually taken by my dad on an early digital camera.
Royle, Stephen: I don't have many pictures in the of me in the lab actually. Like I discovered as looking back. That's kind of, you know, kind of a long time ago when we didn't take cameras everywhere. So that's a Faraday cage. And there's some amplifiers next to me there. So it's definitely electrophysiology lab and together with Banovich, who's on the right in that photo up there, she was a postdoc in the lab. And we were kind of sick as these during this time. I had to. Grateful on. It was absolutely fantastic time doing my PhD.
Royle, Stephen: We together, the two of us, we worked out that these receptors HP gated ion channels, P2X receptors. They were the kind of big thing in the drug industry at the time.
Royle, Stephen: So that there was a lot of interest in these receptors, we realised that two of them feedback four and 2X2 were trafficked differently and this was kind of for neuroscientists, they hadn't really thought about how receptors were moved around inside cells because they didn't really have that cell biology angle. And so really this is where we came in. We we realised that.
Royle, Stephen: To P2X4 was packaged into vesicles, which I'm seeing that image there, and because these receptors control the excitability of the cell, if you remove receptors from the cell surface then you make the cell less excitable and vice versa. And so we realised that membrane traffic was really important for controlling cellular excitability.
Royle, Stephen: So how receptors get moved is shown in this movie here. So the receptors for those blue things on cell surface, and now we're inside the cell. I'm just playing this because it's important for the rest of the talk.
Royle, Stephen: We're gonna see classroom mediated endocytosis here. So classroom is this yellow 3 legged protein that is coming together making a lattice and and pulling this speckicle inwards. And now you can see through the plasma membrane the receptors being pulled in and targeted into this vesicle. So the vesicles forming and there's this classing cage around it. The details of that Cage were worked out by Colin Smith who is here in the audience and this is how a class encoded vesicle is made finally.
Royle, Stephen: It needs to be pinched off. That's what we're going to see. Now with this purple protein here.
Royle, Stephen: And basically this is how you take up a bunch of membrane containing receptors inside the cell and and send it somewhere else for further processing. So this was what I was working on for my PhD and when it came to doing a postdoc, I decided I wanted to keep working on this membrane traffic stuff.
Royle, Stephen: So I, but I wanted to keep working in neuroscience, so one of the big questions at the time was how sign up to vesicles are recycled.
Royle, Stephen: So you have a pre synaptic terminal which has a bunch of synaptic vesicles.
Royle, Stephen: They release neurotransmitter by fusing with the plasma membrane and then the vesicle must be reformed refilt with neurotransmitters so it can signal again. This is how neurotransmission works, so that bit was known. What wasn't known was how the vesicles were retrieved, and in the 1970s there came these two models. So Hoisa Reis felt that it was via classroom mediated endocytosis, which we all know what that is, because I just showed you a movie. And then the other version, I've had a better.
Royle, Stephen: PR it was called Kiss and Run.
Royle, Stephen: Which is a much more memorable term than classroom engagement, the cytosis. And so a lot of people felt that the fescue was just briefly kissed the plasma membrane and then came back away. We thought this just couldn't be true, so we wanted to test the two models we being myself and Leon. Like Nader, who's the gentleman on the left.
Royle, Stephen: If his name sounds familiar, it's because he's the husband of Ruth. Marilla Wagner, who's my PhD supervisor. So that was rather cosy. Perhaps more cosy is the fact that the gentleman next to him is Peter McNaughton, who was his PhD supervisor and he was the supervisor of my wife. Who said today.
Royle, Stephen: So it was. It was all very cosy.
Royle, Stephen: This pictures taken at Leon's fresh shrift a few years ago in Sussex, so I was based here at the LMB, the building where we were is it's just been built here with this. This is my cursor. Yeah, this little crane here. But actually, even if the building was built, you wouldn't have been able to see the lab because it was down in the basement. So it it was a visual neuroscience lab, and we would spend a lot of time in the dark because a lot of the experiments were light sensitive.
Royle, Stephen: And I can tell you in the winter time, cycling to work in the dark, spending all day in the dark and then cycling home in the dark was not fun for months on end. But we had we had a lot of fun in in the lab anyway regardless. So what I wanted to do was to deplete clathrin, so reduce the levels of clathrin in neurons and test the toys erase model, that was that was my goal. And I did do that and we did publish a paper on that and it is my most highest cited paper. But I'm not going to talk about that today because I discovered something else.
Royle, Stephen: Trying to get that working, which ended up launching my independent research career.
Royle, Stephen: So the plan was to use RNAI to deplete flat rate and at the time that was a new technology. And in fact in the original R and AI paper, it says R and AI does not work in neurons. So this was a we felt that maybe it might. So we were getting willing to give it a go. But because of this uncertainty, I wanted to just trial it in non neuronal cells at the same time. And so I started depleting clathrin in regular cell lines.
Royle, Stephen: And discovered that the cells didn't divide normally, so you might think well, that's because you've interfered with this membrane traffic thing. But I also realised this is a repeat of an experiment that I did. This is a repeat done by Fiona Hood, who was the first postdoc in my group.
Royle, Stephen: What I found was that in non dividing cells that's this image here is in these pits and vesicles. So these tiny dots doing its membrane traffic thing. Whereas when the sold starts to divide.
Royle, Stephen: And jumps onto the mitotic spindle. Hopefully you can see that here.
Royle, Stephen: So we've got a cell division problem. When we depleted clathrin and we could see classrooms on spindle and this suggested to us that clustering has a second function besides doing all that membrane traffic stuff during mitosis. And so while I was a postdoc, fortunately, you know, Leon was a visual neuroscientist and this was really not his interest at all. But amazingly, he let me work on this.
Royle, Stephen: And obviously take it away afterwards because he wasn't interested in the in the slightest. But we published a couple of papers on this, and what's these papers sort of hint at is is what I then worked on as I set up my own group at the University of Liverpool. So I'm going to describe what the role of clathrin is in mitotic cells and how it's mitotic function. OK, so the mitotic spindle, which I showed you before is that machine that separates the chromosomes to the two daughter cells.
Royle, Stephen: This is a high voltage EM view of that same thing. This is the spindle here and you can see there are these bundles of microtubules that connect to the chromosomes which are here on the metaphase plate. So it's single microtubule, it's a bundle of microtubules and actually they're held together by stuff. So we've known for a long time that there's some stuff between these microtubules, microtubules. So in the image on the left, the K fibre, the kinetical fibre, which is this bundle which is this bundle is coming towards us and then microtubules are these tiny circles.
Royle, Stephen: So they're 25 nanometres in diameters. This is really, really small, and you can see that between them there's some electron density. If we section the K fibre the other way. So now the microtubules appear as stripes. You can see that there's like these tiny wrongs on the ladder.
Royle, Stephen: And we've, we've thought that clathrin might be doing this connection of the microtubules so you can understand how this might work if you think about a kind of real world example here.
Royle, Stephen: If you have some scaffolding, if a if a if AK-5 was like some scaffolding. If you put just the metal poles up there, that platform right at the top, which would be the kinetical would be very unstable. If you were stood on there, that would be not stable at all. So what scaffolders do and what cells have evolved if a way of cross bracing all of those long struts so that the structure becomes more stable.
Royle, Stephen: And this is the way it seems to work in cells as well. So this is a 3D toomogram made by Dan Booth who was my first PhD student. He's now a runs his own group in University of Nottingham.
Royle, Stephen: And we could see that this this stuff that's between the microtubules is a more complex than just a simple strut. And we thought the clathrin could fulfil this function so he was able to label using an antibody that had a gold particle on it.
Royle, Stephen: See it between the microtubules when we section the cells, so that suggested that clathrin could indeed do this cross bracing function.
Royle, Stephen: And what Fiona Hood did in the lab was to. We realised that classroom probably didn't do this on its own because it can't really bind to microtubules by itself. So what she did is she purified the spindle clathrin complex. So.
Royle, Stephen: She there's some aspect data at the top and then below it you'll see we found these two of the proteins tax free and chapter talk, whose names come from the fact that they're overexpressed in different cancers. So they were in the right place at the right time.
Royle, Stephen: To be doing to be sort of joining hands with clatheryn and and helping it do its function, there were other.
Ngan Lam: Oh yeah.
Royle, Stephen: There were other.
Royle, Stephen: Yeah, no problem there. So there are other.
Royle, Stephen: Reasons why we we we thought we got the right things. They're in the right place at the right time. They had other similarities, but the other thing that confirmed that we got the right thing was three of the labs found the same thing at the same time. So a group in Germany, group in Taiwan and a group in China.
Royle, Stephen: All working in different systems. Mammalian Xenopus identified the same complex, so we knew we got the right thing.
Royle, Stephen: What Fiona was ultimately able to do was to assemble microtubules in the test tube and these are the white lines in the top or the dark things in the bottom and she could incubate with the complex that she purified and get microtubule bundles to form. So compared to PLC one which was a protein that can bundle microtubules on its own, she was able to make microtubule bundles just in the test tube. This is completely outside of the cell.
Royle, Stephen: Suggesting that we really had found the thing that sticks the K fibre together.
Royle, Stephen: So the way we think it works is like this. That platform is part of a team holding this, the microtubules together and we now know quite a lot of details about this. So there's another protein gyutsu, one that has joined the party and they they bind together shown by those dotted lines there. And there's a there's a sort of schematic diagram here which shows that clathrin 1 foot of clathrin sticks to the microtubule and this part of TACK 3 we think sticks the microtubule as well.
Royle, Stephen: Whereas this protein chapter talks binds to TACK 3 but doesn't bind to the microtubule and Gypsy 1 binds to clathrin, it doesn't bind to the microtubule so they're like accessory subunits.
Royle, Stephen: We know the details of this so that the this dotted line here was worked out by Alex Birds Lab in Dortmund, but this interaction here was figured out by our lab together with Richard Bais Group and we we know the structure of this part of TACK 3 that binds to clathrin here.
Royle, Stephen: And the other dotted line here will be published later this year. This is how chapter TOG interacts with TACK 3. It binds to this coil to coil region here, and you might be thinking, oh, this is getting a bit detailed. A bit boring, but really this this is very important because if we know how these proteins interact with each other, we can think about ways we can break them apart. Now if we want to treat cancer, we need to stop cells from dividing. And one way you can do this at the moment is to use microtubule poisons which are.
Royle, Stephen: Stopping cell division, but the problem is is every cell in your body has microtubules and poisoning every microtubule in every cell in your body is not a great idea, so they they have a lot of side effects and those side effects can be so bad that you have to stop the treatment. So if we can target this complex specifically, we will only target the dividing cells because as we know, because I showed you the video, clathrin is doing something else in non dividing cells so we shouldn't interfere with.
Royle, Stephen: Cell division in, you know in clathrin function and other.
Royle, Stephen: Cells. So this isn't just pie in the sky. We have a way to do this. So Richard's lab together with Eileen Kennedy in Athens, GA, have developed a peptide that can compete with this interaction.
Royle, Stephen: It's so permeable so we can deliver it to cells and interfere with that interaction. And James Shelford in my lab together with Richard's lab, developed this aphma, which can compete off this interaction here. So we've got two different ways of breaking apart this complex, and hopefully these can be useful in the future for inhibiting cell division in a in a cancer context. OK, so that talk, that part of the talk that was sort of side A were coming to the end, we're going to flip the record over in a minute, but.
Royle, Stephen: That took us from an observation I made in 2002 to a project which is what was going to publish some papers later this year in 2024. So that was 22 years just condensed down into.
Royle, Stephen: Slides and I skipped over the fact that in 2013 I moved here to University of Warwick.
Royle, Stephen: It was a great best decision I made. It's great colleagues here, great research facilities and and a really vibrant community doing mechanical chemical cell biology which hopefully Gavin is understanding a little bit more now. So the just before while we're on the run out groove before we turn the record over I just want to tell you about my mini mission which is to leave science better than how I found it so.
Royle, Stephen: I'm involved in a number of initiatives such as Open science and trying to accelerate scientific communication through preprints. But one other thing that I did a few years ago was wrote a textbook called the Digital Cell. So I'm just going to briefly tell you about this, and then we'll get back to some cell biology.
Royle, Stephen: So I I was kind of motivated to do this because.
Royle, Stephen: I from comments like this really this is a geneticist.
Royle, Stephen: Dumping on cell biology, saying that the whole field of cell biology is a sewer of unreplicated findings, it's really irked me. As a cell biologist. What irks me most about it was I felt it was actually kind of true. So The thing is, is that soul biology has traditionally been a little bit weak on the quantitative side. It's been quite a qualitative science, and coming from a neuroscience background, I felt that we could improve concentration quite a lot.
Royle, Stephen: I'm not alone in this that my colleagues here in Warwick feel the same way and a lot of us take a quantitative approach to cell biology and take, you know, reproducibility seriously. But I think the field could improve a lot more and stop people saying things like that. So I wrote a post about this on my blog and I kind of felt I might write a series of posts about how you can kind of use computers to improve the rigour that you do your cell biology.
Royle, Stephen: And this is kind of June 2016 when I wrote this and I was contacted by Richard Sever at Cold Spring Harbour Laboratory Press. That's Cold Spring harbour there.
Royle, Stephen: They published a number of books.
Royle, Stephen: Some of which we have in my lab. You, if you do any cloning, you'll know the book on the left, which is known as many artists.
Royle, Stephen: So he, he offered.
Royle, Stephen: The opportunity to turn this into a book and actually that June 2016 and we scheduled a phone call for the day after the EU referendum, which was the most depressing day.
Royle, Stephen: Of my adult life and and he was asking me to do I want to write this textbook and actually, you know, when you become an academic, everyone tells you whatever you do, don't write the textbook it takes, it takes loads of effort. They're right. It does. But it was worth doing. So I I did this. It took a few years, but we published digital selling 2019, and it's been translated into Japanese. And that's the Japanese version there, which I really hope says something about cell biology and not. Steven Royle is an idiot.
Royle, Stephen: Anyway, OK, so we're, we're now gonna flip the record over and talk about membrane traffic, which was on the right hand side of that. Those movies that I showed earlier. So we've we've met this before. This is the kind of endocytosis side.
Royle, Stephen: Cells, eukaryotic cells. Our cells in humans have these different membranes, compartments and.
Royle, Stephen: These compartments great because they isolate different activities into different parts of the cell. So each of these compartments has got different proteins and that means they can do different jobs.
Royle, Stephen: The downside of that means is that if you want to move a protein between those compartments, it's a bit of a challenge because they're separated. So what cells have evolved is a way to package proteins into a vesicle and take it, and then make a vesicle fused on the acceptor membrane.
Royle, Stephen: To deliver a a cargo and we already know about this because we we know that at the plasma membrane you can package receptors into a clathrin coated vesicle and put them inwards.
Royle, Stephen: But this happens between all different membrane compartments inside the cell.
Royle, Stephen: So up until a few years ago, we thought we've got the full complement of transport vesicles. There's a few of them shown here. We've already met this one. This is the clathrin coated vesicle. There's a scale bar here, which is 0 to 200 nanometres, and cluster and coves vesicles are about 100 nanometers in diameter.
Royle, Stephen: But it turns out we didn't know all of these transport vesicles, so we found another one in my lab and I'm just going to briefly tell you about that. So the experiment I'm showing here is a absolutely pivotal experiment, which was done by Gabrielle Laroc in in my group. And what she's doing here is adding a compound to the cells and trapping a protein that we've done some genetic trickery to to make it stick to the mitochondria. And the reason we were doing this experiment.
Royle, Stephen: Was.
Royle, Stephen: And we wanted to kind of inactivate the protein, but actually it turned out to be a way of capturing small vesicles because this protein turned out to be tightly associated with a transport vesicle that had not been discovered previously.
Royle, Stephen: So we got suspicious that there was something going on with this experiment because the mitochondria looked like they were beginning to stick together. And so together with Nick Clark, who was our PM person at the time.
Royle, Stephen: The two of them did these experiments where.
Royle, Stephen: But it's a trap. The protein onto the mitochondria. This is 20 seconds after addition. This is 5 minutes and this is half an hour and you can see it half an hour. The mitochondria are all kind of clumpy. So they process these cells and look to them under the electron microscope. And they could see that. Sure, enough, the mitochondria were stuck together, but they were held together by these tiny vessels. You can see these little circles are studying the surface of the mitochondria.
Royle, Stephen: And these are about 30 nanometers in diameter and we, we call them intracellular nano vesicles or imvs. So what was going on was we were trapping this protein. She's called TPD. It doesn't really matter for this talk, but we're trapping it on the mitochondria. But this protein was very tightly attached to this imv here. And because there's lots of TP DS on the imv.
Royle, Stephen: The mitochondria. We can't stick together.
Royle, Stephen: So that allowed us to find this new transport vesicle, which we can put on this sort of scale diagram as the smallest transport vessel in cells. Now in that movie where you saw those vesicles being relocated to the mitochondria, the the image beforehand was very sort of fuzzy and we couldn't see them very well. But now we've developed ways to look at these vesicles.
Royle, Stephen: A lot better. So we've got better resolution in space and time that allow us to watch these vesicles moving around.
Royle, Stephen: So this movie is actually playing at real time. You can see the the counters going up in seconds and the top left, and there's thousands of these vesicles inside cells. And there's a there's a blow up on the right. So you can have a look and see if you can follow one for a few frames. They they move with mainly diffusively. This is a movie taken by Megan's as well in my lab. So we're we're really intrigued by these vesicles because they're brand new. There's lots to find out about them. We think they've got lots of different sources.
Royle, Stephen: Lots of different destinations and sell. So we think there's lots of different identities and we kind of think of them as like Skittles and we're keen to identify the different flavours of of that we find inside cells. And So what we've been doing most recently is.
Royle, Stephen: Doing some proteomics so we can isolate the vesicles and we can see what proteins they've got associated with them, and we can therefore we can look at the cargos they're transporting and see what what flavours we might have in inside the packet. That is the cell.
Royle, Stephen: So not very important for the this talk at all, but there's lots of different cargoes. So we can see lots of different transporters and receptors and cellodes molecules. So these are all the things that have been moved by the imvs.
Royle, Stephen: And what we're trying to do is unpick the different flavours and the different functions that these imvs might be involved in. So a current project is Mary Fysenko in the lab is identifying a type of vesicle that's involved in autophagy, which is a way that cells used to eat components to recycle the material. So that's an ongoing project. And I'd just like to finish with a another couple of flashy movies to to tell you about what you can do with these.
Royle, Stephen: With this information that we get out cell biology studies. So one thing that we've been doing a lot of in my lab is generating tools for cell biology. And so we can do this by, oh, that's not plain again, is it? Damn it, it's working earlier anyway.
Royle, Stephen: I'll play it again in a minute. When I've told you what it is supposed to be looking at. So if we understand cell biology and how proteins come together to generate a function, it means that we can remix them.
Royle, Stephen: So we can generate new cell biology by taking components and then controlling them ourselves. So what we're doing on the left here is we're generating clutter encoded vesicles just in this region of the cell here. And we're doing that using light. So this is a way that we can take the information that we've learned from doing cell biological studies and and generates a controllable system, in this case using light to enable us to internalise the things that we want just in one region of the cell.
Royle, Stephen: We can also do like weird stuff as well, so we can make class and go to vessels appear in places where they shouldn't. So on the right is a movie made by Chancey Kuy, which is showing classic vesicles being formed on the mitochondria, which is definitely a place where they shouldn't form. But you know, we're the boss, we can make it happen. So this is, you know, it's partly it's not just having fun in the lab. It's about, you know, do we really understand these systems enough to re engineer them and generate new function in places where it hasn't been previously?
Royle, Stephen: To test whether we really understand these systems.
Royle, Stephen: Another example is we re engineered ferritin, which is an iron binding protein because iron is electron dense, we can see it in the EM. So what we what Nick Clark did is he came up with a way to target the ferritin molecule.
Royle, Stephen: Particle to a protein of interest. And so he's he's labelling that this protein here which is in cluster encoded pits.
Royle, Stephen: And we can see the location of the ferritin. Therefore the location of this protein we're interested in and map it out using a computational approach.
Royle, Stephen: And the final example I just want to show is from Laura Downey and my group. She's been looking at membrane contact sites. So this is something kind of related to membrane traffic, but a little bit different. Some of the organelles are very in very close contact and we really lack tools to be able to label those up nicely.
Royle, Stephen: And look at them in different stages of the cell cycle, for example. So this is something that Laura's been working on and she's developed a way to be able to label in this contacts between the endoplasmic reticulum and the mitochondria. And this is a new tool that we're we're just writing paper at the moment and I'm very excited to to get that out into the wider world.
Royle, Stephen: OK, so I just want to finish by thanking a number of people that were involved in all those lovely movies and things that I've been able to show to you today. So there's a list of everyone that's been in that I think it's complete.
Royle, Stephen: To date, the the dark, the ones that are involved are the people currently in the group and so thank you to everyone for over the years for all your hard work and dedication. Following that we have Cam do, which is our computing and advanced microscopy development unit here at the medical school.
Royle, Stephen: Of them all our microscopes and computing needs would be in ruin, and then I'd like to thank collaborators. I mentioned Richard Bayless, who we've done the structural work with Ian prior. We did the early EM work with, but we have ongoing collaborations with a number of people, including Collin Smith and our boys Lambert. And finally, I'd like to thank the people that funded the work without them. Obviously, I couldn't do any of this stuff. So. So thank lots of thanks. It's like the Oscars, isn't it? But anyway, there's finish there and thank you very much for your attention.
Royle, Stephen: Great. Thank you for a fantastic talk. We've got time for a few questions, so.
Royle, Stephen: We'll start in the the the room, and then I'll turn to the colleagues that connected online. So any any questions for Steven?
Royle, Stephen: That's like.
Royle, Stephen: Ha ha.
Royle, Stephen: I think you've described a really interesting journey that's taking you kind of up and down. You know, the length of the country into lots of different, you know, interesting places. If you were to look back across the whole journey.
Royle, Stephen: And you know, give yourself some advice imagining that you were at the beginning again. You know what? What, what would that be?
Royle, Stephen: Yeah, I don't know. It's I've. I've done sort of talks before, you know, kind of career talks for early career researchers from an academic perspective.
Royle, Stephen: And actually, it's quite hard to give sensible advice because that was a long time ago when things things have changed scientists a lot more competitive these days, and there's a lot lots of challenges facing today's career researchers. So. So don't listen to all older people like me, I guess.
Royle, Stephen: Don't take my advice.
Royle, Stephen: Right. We got another question that if you?
Royle, Stephen: Do you mind repeating the question with the people online? The question was, is the digital solution going to be translated in any of the languages? Not not that I know of, yeah.
Royle, Stephen: It would be great, but yeah.
Royle, Stephen: I'm sure you could get one of your clever computers to do it with AI.
Royle, Stephen: Any other questions in the room?
Royle, Stephen: Have we had any come in on life?
Royle, Stephen: He just wants. Hopefully it's not sorry, I've got to go now. Yeah. Just wanna say yeah.
Royle, Stephen: It.
Royle, Stephen: You create them, which is hard questions, isn't it?
Royle, Stephen: No. Well, no. So so that I think is a green foraging. The carrier carrier means body and you means.
Royle, Stephen: Thank you.
Royle, Stephen: So. So yeah, a lot of them have got classical derivations clathering that I talked about today gets its name, but the flat rate means lattice like in Greek and proteins. Turns out in on the own. So that's how typically a protein will be named plastically. But yeah, if you discover these things, unfortunately.
Royle, Stephen: A lot of that's been done now, so there's no not much naming of new things these days.
Royle, Stephen: Thank you for the question. Yeah.
Royle, Stephen: Not gonna actually. Umm. So what, you're working on this the where they come, where they go, do they come address existing funding. Do you think they want to have? Yeah. So this is something we're really actively working on at the moment and we there's there's a few hypothesis one is that there's a common mechanism of formation because and that's hinted up by the fact that there are quite a uniform size so you kind of think there might be a cookie cutter type of mechanism to make them that's common to all of them but then we do have all these different flavours so maybe there are different ways of formation this is a long winded way of saying we don't know and we're working on it but yeah.
Royle, Stephen: Yeah, I think I I think so. I think a lot of people that work on festival transport focus on more interesting cells that have got long processes where the transport is, you know, kind of has to be engineered to be, you know, for the vessel to get to its destination. But most of the transport that needs to happen is very short distance, where diffusion is just able to do the job. So yeah, I think a lot of the time the the distance is quite proximal and diffusion's fine.
Royle, Stephen: Any other questions in the room? I've managed to work the microphone out now, which is clearly good.
Royle, Stephen: Hold on. Well, would you like to guess how many kinds of land these are?
Royle, Stephen: Yeah, I think a lot. No, I see.
Royle, Stephen: I think it all depends on your definition and how you're classifying. Obviously and like how I mean, I guess at some level they're all different because they all have different numbers of cargo molecules in different complements.
Royle, Stephen: But stepping back a bit, we know already that there are 16 different rabs on these less goals out of 44 that we've tested.
Royle, Stephen: So the minimal case would be 16, say, but it could be could be higher.
Royle, Stephen: We've got another question over there, so.
Royle, Stephen: The set block of independent create do you think Catherine has any knowledge?
Royle, Stephen: And again, if you can repeat the question, yeah, sorry. So the question was, does classifying have other roles? It's got a role in member in traffic and a role in mitosis. But does it have other roles? I mean, there have been some, I actually I went to a meeting when I first did the mitotic clathrin stuff, and I had a poster at this meeting in Madrid and there was a another scientist there who got another poster saying classroom was involved in transcription. And I thought I had this sinking feeling like, oh, no, is this what happens? Like there's these crazy people who think that.
Royle, Stephen: Our teams do new functions and it's all not true. Nothing ever happened about the transcription thing that was proposed by by another group, but it didn't really go anywhere, so I don't know. Never rule it out. But yeah, don't I think 2's enough for any protein.
Royle, Stephen: Right. So I'm not seeing any other questions in the room and I think that the question that was coming in online was when when can you share the recordings?
Royle, Stephen: Go back to them. But I'd just like to, you know, wrap things up by again, you know, congratulating you on your achievements. It was fantastic, you know, hearing about the breadth and the depth of the work and to say how delighted we are to have you as a member of our faculty here at the medical school. Thank you very much.