Cancer Research UK Director of Research in Cell Division
Departments and Institutes
Dr Pine's research focuses on the detailed understanding of cell division (mitosis) and the interplay between protein kinases, protein phosphatases, and APC/C-mediated proteolysis. This interplay regulates the entry to mitosis and the coordinate chromosome segregation with cell separation to ensure that the two daughter cells receive an equal and identical copy of the genome.
Since mitosis is a highly dynamic process we study living cells by time-lapse fluorescence microscopy, but complement this with biochemical analyses on cells in which we have knocked-out or mutated specific mitotic regulators using somatic cell recombination.
We developed a FRET biosensor to assay Cyclin B1-Cdk1 kinease activity in vivo and are using this to define the pathways that regulate the timing of mitosis. SILAC mass spectrometry is being used to analyse protein complexes through the cell cycle and to identify the proteins responsible for regulating the Cyclin-Cdks.
To understand how proteolysis regulates progress through mitosis we complement the analysis of APC/C-dependent degradation in living cells with biochemical analyses of protein complexes and ubiquitination activity. These studies are revealing how the APC/C is activated and how it is able to select a particular protein for destruction at a specific time. Moreover, the intimate coupling of the APC/C with the spindle assembly checkpoint that is essential to the control of chromosome segregation has meant that our recent work has begun to elucidate the key events in the checkpoint pathway.