Quantitative Phosphoproteomics Reveals the Signaling Dynamics of Cell-Cycle Kinases in the Fission Yeast Schizosaccharomyces pombe

Abstract

Summary: Multiple protein kinases regulate cell-cycle progression, of which the cyclin-dependent kinases (CDKs) are thought to act as upstream master regulators. We have used quantitative phosphoproteomics to analyze the fission yeast cell cycle at sufficiently high temporal resolution to distinguish fine-grain differences in substrate phosphorylation dynamics on a proteome-wide scale. This dataset provides a useful resource for investigating the regulatory dynamics of cell-cycle kinases and their substrates. For example, our analysis indicates that the substrates of different mitotic kinases (CDK, NIMA-related, Polo-like, and Aurora) are phosphorylated in sequential, kinase-specific waves during mitosis. Phosphoproteomics analysis after chemical-genetic manipulation of CDK activity suggests that the timing of these waves is established by the differential dependency of the downstream kinases on upstream CDK. We have also examined the temporal organization of phosphorylation during G1/S, as well as the coordination between the NDR-related kinase Orb6, which controls polarized growth, and other cell-cycle kinases. : Swaffer et al. use phosphoproteomics at high temporal resolution to determine the fine-grain differences in substrate phosphorylation timing during the fission yeast cell cycle. This global analysis reveals how multiple different cell-cycle kinases contribute to phosphorylation ordering, as well as the hierarchy of mitotic kinases downstream of the master regulator CDK. Keywords: cell cycle, mitosis, CDK, cyclin-dependent kinase, cell-cycle kinases, kinase networks, protein phosphorylation, phosphoproteomics, fission yeast, Schizosaccharomyces pombe