Live-cell imaging defines a threshold in CDK activity at the G2/M transition Hironori Sugiyama, Yuhei Goto, Yohei Kondo, Damien Coudreuse and Kazuhiro Aoki Developmental Cell 10.1016/j.devcel.2023.12.014



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Cyclin-dependent kinase (CDK) determines the temporal ordering of the cell cycle phases. However, despite significant progress in studying regulators of CDK and phosphorylation patterns of CDK substrates at the population level, it remains elusive how CDK regulators coordinately affect CDK activity at the single-cell level and how CDK controls the temporal order of cell cycle events. Here, we elucidate the dynamics of CDK ac- tivity in fission yeast and mammalian cells by developing a CDK activity biosensor, Eevee-spCDK. We find that although CDK activity does not necessarily correlate with cyclin levels, it converges to the same level around mitotic onset in several mutant backgrounds, including pom1D cells and wee1 or cdc25 overexpress- ing cells. These data provide direct evidence that cells enter the M phase when CDK activity reaches a high threshold, consistent with the quantitative model of cell cycle progression in fission yeast.