About the Project
University of Manchester Supervisor: Dr Christoph Ballestrem, A*STAR Supervisor: Edward Manser (IMCB)
Project description: Cells continuously sense and remodel their extracellular environment. We are interested in the mechanisms underpinning this sensing and remodelling, so as to find new ways to prevent disease and help regeneration.
The main devices cells use to sense and remodel the extracellular matrix (ECM) are focal adhesions (FA). These FAs contain adhesion receptors (integrins) that, when activated, associate with the intracellular actin cytoskeleton via a series of protein intermediaries (perhaps 40-50 components) that form a dynamic macromolecular machine. This machine can exert force on the ECM and therefore move both the cell and the substrate. The beta-integrin (cytoplasmic domains) can bind a very a limited number of these proteins directly in vivo and it is generally considered Talin, Kindlin and Tensin are the most important (each of these has 2 or 3 human isoforms). These adaptor proteins allow for additional recruitment of structural and regulatory components such a protein kinases (some of which are oncogenes). To date the Tensins are the least studied of the important integrin-binders and we do not understand their contribution to assembly the FA network, particularly ’fibrillar adhesions’.
We are particularly interested in how cells transduce integrin-mediated signals derived from the ECM. Here we put forward an interdisciplinary collaborative proposal between the Ballestrem lab at Manchester (University of Manchester) and the Manser lab in Singapore (A*STAR) to investigate how a network of adhesion regulatory proteins (Tensin 1, 2 and 3) controls fibrillar adhesions that are particularly important for remodelling and the biophysical properties of the extracellular matrix. Because Tensin-mediated formation of adhesions is known to be regulated by protein kinases such as AMPK (Georgiadou, et al., 2017) the student will also have the opportunity to investigate the role of Rho-associated kinases ROCK, PAK and MRCK (Zhao & Manser, 2015) in these processes.
Aberrant regulation of the focal adhesion complex underlies several disease states including age-related tissue degeneration, cancer, fibrosis and cardiovascular disease. The project will comprise a wide variety of methods to gain insight into the mechanisms of how this protein works in normal and cancerous cells. Methods include state-of-the-art imaging techniques and sophisticated high throughput unbiased mass spectroscopy techniques.
Methods: This project is highly interdisciplinary and involves a large number of methods that span areas of cell biology: advanced fluorescence microscopy (super-resolution; FRET; FCS); molecular biology (cloning; RNAi; CRISPR-Cas9); biochemistry and mass spectrometry
Funding Notes
This project is available to UK/EU candidates. Funding covers fees and stipend for four years. Overseas candidates can apply providing they can pay the difference in fees and are from an eligible country. Candidates will be required to split their time between Manchester and Singapore, as outlined on www.manchester.ac.uk/singaporeastar.
Applications should be submitted online and candidates should make direct contact with the Manchester supervisor to discuss their application directly. Applicants must have obtained, or be about to obtain, at least an upper second class honours degree (or equivalent) in a relevant subject.
References
1. Stutchbury B, Atherton P., Tsang R., Wang D-Y, and Ballestrem C. (2017) Distinct focal adhesion protein modules control different aspects of mechanotransduction. Journal of Cell Science. doi: 10.1242/jcs.195362.
2. Atherton, P., Stutchbury, B., Wang, D., Jethwa, D., Tsang, R., Meiler-Rodrigues, E., Wang, P., Bate, N., Zent, R., Barsukov, I.L., Goult, B.T., Critchley, D.R. and Ballestrem, C. (2015) Vinculin controls talin engagement with the actomyosin machinery. Nature Communications. 6:10038.
3. Roux, K.J., Kim, D.I., Raida, M., and Burke, B. (2012). A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. The Journal of Cell biology 196, 801-810.
4. Dong, J.M Tay, F.P Swa, H.L. 2 Gunaratne, J. Leung, T. Burke, B. and Manser, E. (2016)
Proximity biotinylation provides insight into the molecular composition of focal adhesions at the nanometer scale Sci Signal. 2016 Jun 14;9(432):rs4.