KINETOCHECK · Dissecting the signalling cascade in the vertebrate spindle assembly checkpoint
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2004-03-01 → 2006-02-28
- EU contribution
- €214,590
- Participants
- 1
- Scheme
- EIF
Lines connect the coordinator with its partners.
Results in brief
Final Activity Report Summary - KINETOCHECK (Dissecting the signalling cascade in the vertebrate spindle assembly checkpoint)
Accurate chromosome separation is highly controlled by a checkpoint, because an error during this step cannot be corrected. If this checkpoint is deficient, cells with abnormal chromosome number will be produced, which is a hallmark of cancer. In order to study this process, we studied 3 proteins situated at the anchoring point of the chromosome to the separation machinery, a beautiful cellular structure called the mitotic spindle. I studied how CENP-E is properly localised, and I found by which domain it binds to its partner BubR1, which is strictly correlated with its proper localization. This suggests that direct binding of CENP-E to BubR1 is required for the correct localisation of CENP-E. I have also identified several modifications (by phosphorylation) of CENP-E and Mps1, and I am currently pursuing my work to understand the mechanistic details of CENP-E regulation by Mps1.
Data: CORDIS, © European Union
Project objective
The mitotic checkpoint acts to inhibit entry into anaphase until all chromosomes havesuccessfully attached to spindle microtubules. Unattached kinetochores are believed to release inactivated form of proteins such as BubR1 and Mad2 that inhibits APC/C-dependent ubiquitinylation and subsequent proteolysis of components needed for anaphase onset.Previous work by Dr. Abrieu(the host laboratory) has shown that the kinase Mps1 is required to recruit some checkpoint components onto the kinetochore, including CENP-E, a kinesin-like protein acting as a sensor of kinetochore to microtubules attachment. My work in this lab will consist on further dissecting the regulation of CENP-E by Mps1 in this context, using biochemical, chimiogenetic and microspectroscopic approaches. This will involve developing a new method to identify Mps1substrates; a multidisciplinary approach (chimiogenetic) in tight collaboration with the organic chemistry laboratory of Prof. Perigaud. I am also proposing to identify the minimal kinetochore binding domain of CENP-E. This will allow me to follow its activation, and regulation of the downstream kinaseBubR1 at the kinetochore by FRET (Fluorescence Resonance Energy Transfer). Finally, I will try to uncover new kinetochore proteins by fishing for binding partners of two outermost kinetochore components (CENP-E and Mps1) and the innermost centromeric protein (CENP-A). Altogether, this proposal should provide a better understanding of the checkpoint regulation at the kinetochore, a process whose deregulation is leading to aneuploidy, a hallmark of cancer.Hopefully, these studies could lead to new therapeutic development specifically targeting cancer cells by directly activating checkpoint regulators, thus preventing side effects onto normal cells.
Original text from CORDIS.
Participants
- CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE · PARISCoordinatorFrance
Links
Data: CORDIS, © European Union