Olafsson, Gudjon;
(2018)
Manipulating kinetochore function with synthetic physical interactions.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
The kinetochore is a large protein complex that assembles on centromeres. It enables accurate chromosome segregation by attaching chromosomes to the mitotic spindle via microtubules and by recruiting regulators that control mitosis. Many of these regulators modify kinetochore components by post-translational modifications such as phosphorylation and ubiquitination. For example, when kinetochores fail to attach to microtubules, the kinase Mps1 phosphorylates the kinetochore to activate the spindle assembly checkpoint (SAC). The abundance of chromosomal instability mutants suggests that numerous kinetochore regulators remain to be identified and the mechanisms of some known regulators are poorly characterised. The kinetochore is highly conserved from yeast to humans and many important discoveries of kinetochore function have been made using Saccharomyces cerevisiae. In order to identify kinetochore regulators in budding yeast, I have used the Synthetic Physical Interaction (SPI) technology to separately associate each member of the yeast proteome with a kinetochore protein. In addition, I have recruited several hundred proteins involved in chromosome segregation and post-translational modifications individually to kinetochore proteins representing each of the major kinetochore subcomplexes. I then associated each candidate regulator separately with most of the kinetochore proteins to spatially map their function. This method identifies candidate kinetochore regulators such as chromatin remodelling complexes, phosphatases and kinases; some of which are novel. In this thesis, I focus on three regulators; first, the Cdc14 phosphatase, second, components of the SAC and lastly the Polo-like kinase. These studies reveal kinetochore proteins sensitive to specific regulators and that their action is spatially restricted within the large kinetochore structure.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Manipulating kinetochore function with synthetic physical interactions |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| UCL classification: | UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10041050 |
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