Understanding mitosis using real time analysis of mitotic protein kinase activity

We have a developed a probe to look in living cells at the activity of the factor that causes cells to divide. We will use this probe to measure when this activity first appears and then alter the cell in defined ways to work out what causes it to become active. This work should help us to understand how cells trigger cell division and eventually may lead to a better understanding of how this can go wrong in cancer.

We have developed a FRET biosensor that specifically measures Cyclin B1-Cdk1 protein kinase activity in living cells. We intend to use this biosensor to understand how cells regulate the entry to mitosis. We will measure when and where Cyclin B1-Cdk1 is activated as unperturbed cells enter mitosis, and then assay how this is affected when we deplete or overexpress specific proteins that have previously been implicated, directly or indirectly, in the control of mitosis. In this way we should be able to define more exactly how Cyclin B1-Cdk1 is activated at mitosis.