About the Project
Background
For chromosome segregation into daughter cells to occur without error, mammalian cells need to round up and adopt a spherical shape during mitosis. If rounding does not take place, e.g. in dense stromal tissue, mistakes are made that result in aneuploidy, multiploidy and aberrant growth. There is evidence that cells stop dividing if the extracellular conditions are inappropriate and that the G2/M cell cycle checkpoint receives signals from the cell microenvironment. Currently, the identity of these signals, which link adhesion receptors and their associated complexes to the cell cycle machinery, is unknown.
Aims and approaches
To identify these signals, the supervisory team has established methods to isolate adhesion complexes (1-3) or tag receptor-associated proteins using proximity biotinylation (4), define them by quantitative protein or phosphoprotein mass spectrometry (1-3), and probe the functional roles of candidate proteins/post-translational modifications using bioinformatics, protein biochemistry and high-end light microscopy (1,5,6). All techniques are in use in the Humphries lab, with the Ballestrem lab providing particular expertise in high-end light microscopy.
Questions and outcomes
Potential questions to be addressed in the project include: (a) the identity of the links between adhesion receptors and the cell cycle machinery that are responsible for cell rounding during mitosis, (b) the cell cycle-dependent changes that take place in adhesion complexes, and (c) the mechanisms whereby extracellular matrix rigidity is converted into signals that control cell proliferation. Insights into any of these processes will improve our understanding of cell replication in a range of conditions and suggest ways to control proliferation.
http://www.humphrieslab.manchester.ac.uk
http://www.wellcome-matrix.org/research_groups/martin-humphries.html
https://www.ballestremlab.com/
http://www.wellcome-matrix.org/research_groups/christoph-ballestrem.html
References
1. Byron, A., Askari, J.A., Humphries, J.D., Jacquemet, G., Koper, E.J., Warwood, S., Choi, C.K., Stroud, M.J., Chen, C.S., Knight, D. and Humphries, M.J. (2015) A proteomic approach reveals integrin activation state-dependent control of microtubule cortical targeting. Nature Comm. 6: 6135
2. Robertson, J., Jacquemet, G., Byron, A., Jones, M.C., Warwood, S., Selley, J.N., Knight, D., Humphries, J.D. and Humphries, M.J. (2015) Defining the phospho-adhesome: phosphoproteomic analysis of integrin signalling. Nature Comm. 6: 6265
3. Horton, E.R., Byron, A., Askari, J.A., Ng, D.H.J., Millon-Frémillon, A., Robertson, J., Koper, E.J., Paul, N.R., Warwood, S., Knight, D., Humphries, J.D. and Humphries, M.J. (2015) Definition of a consensus integrin adhesome and analysis of its dynamics during adhesion complex assembly and disassembly. Nature Cell Biol. 17: 1577-1587
4. 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. J. Cell Biol. 196: 801
5. Atherton, P., Stutchbury, B., Wang, D.Y., Jethwa, D., Tsang, R., Meiler-Rodriguez, 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 Comm. 6: 10038
6. Stutchbury, B., Atherton, P., Tsang, R., Wang, D.Y. and Ballestrem, C. (2017) Distinct focal adhesion protein modules control different aspects of mechanotransduction. J. Cell Sci. 130: 1612-1624
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