Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS
Ly, T., Whigham, A., Clarke, R., Brenes-Murillo, A.J., Estes, B., Madhessian, D., Lundberg, E., Wadsworth, P., Lamond, A.I.
Intracellular immunostaining, FACS, and proteomics have been combined in a single workflow (PRIMMUS) to isolate and analyse cell subsets defined by intracellular markers of cell cycle progression, thus avoiding potential artefacts associated with arrest-based synchronisation. Key findings include the identification of a class of phosphorylations called ‘early risers’ that increase during G2 phase, and ~100 proteins showing decreased abundances during early mitosis, including RRM2.
This work was completed while Tony Ly was working as a postdoctoral researcher in Angus Lamond's group at the Centre for Gene Regulation and Expression, University of Dundee.