Projects per year
Abstract / Description of output
Chemical-induced dimerisation (CID) uses small molecules to control specific protein-protein interactions. Here, we employ CID dependent on the plant hormone abscisic acid (ABA) to reconstitute spindle checkpoint signalling in fission yeast. The spindle checkpoint signal usually originates at unattached or inappropriately attached kinetochores. These are complex, multi-protein structures with several important functions. To bypass kinetochore complexity, we take a reductionist approach to study checkpoint signalling. We generate a synthetic checkpoint arrest ectopically by inducing hetero-dimerisation of the checkpoint proteins Mph1Mps1 and Spc7KNL1 These proteins are engineered such that they can't localise to kinetochores, and only form a complex in the presence of ABA. Using this novel assay we are able to checkpoint arrest a synchronous population of cells within 30 minutes of ABA addition. This assay allows for detailed genetic dissection of checkpoint activation and importantly it also provides a valuable tool for studying checkpoint silencing. To analyse silencing of the checkpoint and the ensuing mitotic exit, we simply wash-out the ABA from arrested cells. We show here that silencing is critically-dependent on PP1Dis2 recruitment to Mph1Mps1-Spc7KNL1 signalling platforms.
Original language | English |
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Article number | jcs.219766 |
Journal | Journal of Cell Science |
Volume | 132 |
Issue number | 4 |
Early online date | 20 Sept 2018 |
DOIs | |
Publication status | Published - 4 Oct 2018 |
Keywords / Materials (for Non-textual outputs)
- Mps1
- Checkpoint
- Dimerisation
- Mitosis
- Reconstitution
- Spindle
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Dive into the research topics of 'Regulated reconstitution of spindle checkpoint arrest and silencing through chemically induced dimerisation, in vivo'. Together they form a unique fingerprint.Projects
- 4 Finished
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Wellcome Trust Four-Year Studentship (Sadhbh Soper Chafraidh)
Beggs, J.
1/10/14 → 30/09/18
Project: Research
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Profiles
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Kevin Hardwick
- School of Biological Sciences - Personal Chair in Molecular Genetics
- Centre for Engineering Biology
Person: Academic: Research Active