Phosphorylation-dependent binding of mitotic cyclins to Cdc6 contributes to DNA replication control
- PMID: 15496876
- DOI: 10.1038/nature03024
Phosphorylation-dependent binding of mitotic cyclins to Cdc6 contributes to DNA replication control
Abstract
Cyclin-dependent kinases (CDKs) limit the activation of DNA replication origins to once per cell cycle by preventing the assembly of pre-replicative complexes (pre-RCs) during S, G2 and M phases of the cell cycle in the budding yeast Saccharomyces cerevisiae. CDKs inhibit each pre-RC component (ORC, Cdc6, Cdt1/Mcm2-7) by different mechanisms. We show here that the mitotic CDK, Clb2/Cdc28, binds tightly to an amino-terminal domain (NTD) of Cdc6, and that Cdc6 in this complex is unable to assemble pre-RCs. We present evidence indicating that this Clb2-dependent mechanism contributes to preventing re-replication in vivo. CDK interaction with the NTD of Cdc6 is mediated by the cyclin subunit Clb2, and could be reconstituted with recombinant Clb2 protein and synthetic NTD peptides. Tight Clb2 binding occurred only when the NTD was phosphorylated on CDK consensus sites. Human CDKs containing cyclins A, B and E also bound specifically to phospho-NTD peptides. We propose that direct binding of cyclins to phosphopeptide motifs may be a widespread phenomenon contributing to the targeting of CDKs to substrates.
Similar articles
-
Cyclin specificity in the phosphorylation of cyclin-dependent kinase substrates.Nature. 2005 Mar 3;434(7029):104-8. doi: 10.1038/nature03329. Nature. 2005. PMID: 15744308
-
Identification of Clb2 residues required for Swe1 regulation of Clb2-Cdc28 in Saccharomyces cerevisiae.Genetics. 2008 Jun;179(2):863-74. doi: 10.1534/genetics.108.086611. Genetics. 2008. PMID: 18558651 Free PMC article.
-
Cdc6 cooperates with Sic1 and Hct1 to inactivate mitotic cyclin-dependent kinases.Nature. 2001 Jul 19;412(6844):355-8. doi: 10.1038/35085610. Nature. 2001. PMID: 11460169
-
Linking mitosis with S-phase: Cdc6 at play.Cell Cycle. 2008 Mar 1;7(5):597-601. doi: 10.4161/cc.7.5.5519. Epub 2008 Jan 3. Cell Cycle. 2008. PMID: 18256547 Review.
-
Regulation of Cdc28 cyclin-dependent protein kinase activity during the cell cycle of the yeast Saccharomyces cerevisiae.Microbiol Mol Biol Rev. 1998 Dec;62(4):1191-243. doi: 10.1128/MMBR.62.4.1191-1243.1998. Microbiol Mol Biol Rev. 1998. PMID: 9841670 Free PMC article. Review.
Cited by
-
Evolutionary conservation of the CDK targets in eukaryotic DNA replication initiation.Chromosoma. 2015 Sep;124(3):309-21. doi: 10.1007/s00412-014-0500-y. Epub 2015 Jan 11. Chromosoma. 2015. PMID: 25575982 Review.
-
Distinct action of the retinoblastoma pathway on the DNA replication machinery defines specific roles for cyclin-dependent kinase complexes in prereplication complex assembly and S-phase progression.Mol Cell Biol. 2006 Oct;26(20):7667-81. doi: 10.1128/MCB.00045-06. Epub 2006 Aug 14. Mol Cell Biol. 2006. Retraction in: Mol Cell Biol. 2020 Aug 14;40(17):e00319-20. doi: 10.1128/MCB.00319-20. PMID: 16908528 Free PMC article. Retracted.
-
A CDK-catalysed regulatory phosphorylation for formation of the DNA replication complex Sld2-Dpb11.EMBO J. 2006 May 3;25(9):1987-96. doi: 10.1038/sj.emboj.7601075. Epub 2006 Apr 13. EMBO J. 2006. PMID: 16619031 Free PMC article.
-
Regulating DNA replication in eukarya.Cold Spring Harb Perspect Biol. 2013 Sep 1;5(9):a012930. doi: 10.1101/cshperspect.a012930. Cold Spring Harb Perspect Biol. 2013. PMID: 23838438 Free PMC article. Review.
-
Behavior of replication origins in Eukaryota - spatio-temporal dynamics of licensing and firing.Cell Cycle. 2015;14(14):2251-64. doi: 10.1080/15384101.2015.1056421. Epub 2015 Jun 1. Cell Cycle. 2015. PMID: 26030591 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases
Miscellaneous
