title: An integrated chemical biology approach provides insight into Cdk2 functional redundancy and inhibitor sensitivity.
abstract: Cdk2 promotes DNA replication and is a promising cancer therapeutic target, but its functions appear redundant with Cdk1, an essential Cdk affected by most Cdk2 inhibitors. Here, we present an integrated multidisciplinary approach to address Cdk redundancy. Mathematical modeling of enzymology data predicted conditions allowing selective chemical Cdk2 inhibition. Together with experiments in Xenopus egg extracts, this supports a rate-limiting role for Cdk2 in DNA replication. To confirm this we designed inhibitor-resistant (ir)-Cdk2 mutants using a novel bioinformatics approach. Bypassing inhibition with ir-Cdk2 or with Cdk1 shows that Cdk2 is rate-limiting for replication in this system because Cdk1 is insufficiently active. Additionally, crystal structures and kinetics reveal alternative binding modes of Cdk1-selective and Cdk2-selective inhibitors and mechanisms of Cdk2 inhibitor resistance. Our approach thus provides insight into structure, functions, and biochemistry of a cyclin-dependent kinase.
authors.affiliation: Université Montpellier I, 34000 Montpellier, France.
authors.forename: Aude
authors.initials: A
authors.lastname: Echalier
authors.forename: Emilie
authors.initials: E
authors.lastname: Cot
authors.forename: Alain
authors.initials: A
authors.lastname: Camasses
authors.forename: Elsie
authors.initials: E
authors.lastname: Hodimont
authors.forename: François
authors.initials: F
authors.lastname: Hoh
authors.forename: Philippe
authors.initials: P
authors.lastname: Jay
authors.forename: Felix
authors.initials: F
authors.lastname: Sheinerman
authors.forename: Liliana
authors.initials: L
authors.lastname: Krasinska
authors.forename: Daniel
authors.initials: D
authors.lastname: Fisher
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D018360:Crystallography, X-Ray; D051357:Cyclin-Dependent Kinase 2 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D006801:Humans; D007399:Interphase; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010063:Ovum / Q000378:metabolism; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014981:Xenopus / Q000254:growth & development / Q000378:metabolism
chemical_list: D016213:Cyclins; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D016203:CDC2 Protein Kinase; D051357:Cyclin-Dependent Kinase 2