title: Switching Cdk2 on or off with small molecules to reveal requirements in human cell proliferation.
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.
authors.affiliation: Department of Structural and Chemical Biology, Mount Sinai School of Medicine, One Gustave L Levy Place, New York, NY 10029, USA.
authors.forename: Karl A
authors.initials: KA
authors.lastname: Merrick
authors.forename: Lara
authors.initials: L
authors.lastname: Wohlbold
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Jasmina J
authors.initials: JJ
authors.lastname: Allen
authors.forename: Dai
authors.initials: D
authors.lastname: Horiuchi
authors.forename: Noelle E
authors.initials: NE
authors.lastname: Huskey
authors.forename: Andrei
authors.initials: A
authors.lastname: Goga
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
authors.forename: Robert P
authors.initials: RP
authors.lastname: Fisher
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D049109:Cell Proliferation; D051357:Cyclin-Dependent Kinase 2 / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D016193:G1 Phase / Q000187:drug effects / Q000502:physiology; D006801:Humans; D017434:Protein Structure, Tertiary; D016196:S Phase / Q000187:drug effects / Q000502:physiology
chemical_list: C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D000225:Adenine