Switching Cdk2 On or Off with Small Molecules to Reveal Requirements in Human Cell Proliferation

Karl A. Merrick, Lara Wohlbold, Chao Zhang, Jasmina J. Allen, Dai Horiuchi, Noelle E. Huskey, Andrei Goga, Kevan M. Shokat, Robert P. Fisher*

*Corresponding author for this work

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

Multiple cyclin-dependent kinases (CDKs) control eukaryotic cell division, but assigning specific functions to individual CDKs remains a challenge. During the mammalian cell cycle, Cdk2 forms active complexes before Cdk1, but lack of Cdk2 protein does not block cell-cycle progression. To detect requirements and define functions for Cdk2 activity in human cells when normal expression levels are preserved, and nonphysiologic compensation by other CDKs is prevented, we replaced the wild-type kinase with a version sensitized to specific inhibition by bulky adenine analogs. The sensitizing mutation also impaired a noncatalytic function of Cdk2 in restricting assembly of cyclin A with Cdk1, but this defect could be corrected by both inhibitory and noninhibitory analogs. This allowed either chemical rescue or selective antagonism of Cdk2 activity in vivo, to uncover a requirement in cell proliferation, and nonredundant, rate-limiting roles in restriction point passage and S phase entry.

Original languageEnglish (US)
Pages (from-to)624-636
Number of pages13
JournalMolecular cell
Volume42
Issue number5
DOIs
StatePublished - Jun 10 2011

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Merrick, K. A., Wohlbold, L., Zhang, C., Allen, J. J., Horiuchi, D., Huskey, N. E., Goga, A., Shokat, K. M., & Fisher, R. P. (2011). Switching Cdk2 On or Off with Small Molecules to Reveal Requirements in Human Cell Proliferation. Molecular cell, 42(5), 624-636. https://doi.org/10.1016/j.molcel.2011.03.031