XB-ART-15405
J Cell Biol
January 26, 1998;
140
(2):
271-81.
Identification of a preinitiation step in DNA replication that is independent of origin recognition complex and cdc6, but dependent on cdk2.
Hua XH
,
Newport J
.
Abstract
Before initiation of DNA replication, origin recognition complex (ORC) proteins, cdc6, and minichromosome maintenance (MCM) proteins bind to chromatin sequentially and form preinitiation complexes. Using Xenopus laevis egg extracts, we find that after the formation of these complexes and before initiation of DNA replication, cdc6 is rapidly removed from chromatin, possibly degraded by a cdk2-activated, ubiquitin-dependent proteolytic pathway. If this displacement is inhibited, DNA replication fails to initiate. We also find that after assembly of MCM proteins into preinitiation complexes, removal of the ORC from DNA does not block the subsequent initiation of replication. Importantly, under conditions in which both ORC and cdc6 protein are absent from preinitiation complexes, DNA replication is still dependent on cdk2 activity. Therefore, the final steps in the process leading to initiation of DNA replication during S phase of the cell cycle are independent of ORC and cdc6 proteins, but dependent on cdk2 activity.
PubMed ID: 9442103
PMC ID: PMC2132576
Article link:
Grant support: [+]
GM 44656 NIGMS NIH HHS
Species referenced: Xenopus
Genes referenced: cdc6 cdk2 mmut
Article Images: [+] show captions
References [+] :
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Yan, Mcm2 and Mcm3, two proteins important for ARS activity, are related in structure and function. 1991, Pubmed
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Zwerschke, The Saccharomyces cerevisiae CDC6 gene is transcribed at late mitosis and encodes a ATP/GTPase controlling S phase initiation. 1994, Pubmed
Broek, Involvement of p34cdc2 in establishing the dependency of S phase on mitosis. 1991, Pubmed
Bueno, Dual functions of CDC6: a yeast protein required for DNA replication also inhibits nuclear division. 1992, Pubmed
Carpenter, Role for a Xenopus Orc2-related protein in controlling DNA replication. 1996, Pubmed , Xenbase
Chau, A multiubiquitin chain is confined to specific lysine in a targeted short-lived protein. 1989, Pubmed
Chong, Purification of an MCM-containing complex as a component of the DNA replication licensing system. 1995, Pubmed , Xenbase
Cocker, An essential role for the Cdc6 protein in forming the pre-replicative complexes of budding yeast. 1996, Pubmed
Coleman, The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts. 1996, Pubmed , Xenbase
Correa-Bordes, p25rum1 orders S phase and mitosis by acting as an inhibitor of the p34cdc2 mitotic kinase. 1996, Pubmed
Coué, Chromotin binding, nuclear localization and phosphorylation of Xenopus cdc21 are cell-cycle dependent and associated with the control of initiation of DNA replication. 1996, Pubmed , Xenbase
Dahmann, S-phase-promoting cyclin-dependent kinases prevent re-replication by inhibiting the transition of replication origins to a pre-replicative state. 1996, Pubmed
Dalton, Cell cycle-regulated nuclear import and export of Cdc47, a protein essential for initiation of DNA replication in budding yeast. 1995, Pubmed
Diffley, Once and only once upon a time: specifying and regulating origins of DNA replication in eukaryotic cells. 1996, Pubmed
Diffley, Two steps in the assembly of complexes at yeast replication origins in vivo. 1994, Pubmed , Xenbase
Donovan, Replication origins in eukaroytes. 1996, Pubmed , Xenbase
Dowell, Interaction of Dbf4, the Cdc7 protein kinase regulatory subunit, with yeast replication origins in vivo. 1994, Pubmed
Fang, Evidence that the G1-S and G2-M transitions are controlled by different cdc2 proteins in higher eukaryotes. 1991, Pubmed , Xenbase
Fox, The origin recognition complex has essential functions in transcriptional silencing and chromosomal replication. 1995, Pubmed
Gavin, Conserved initiator proteins in eukaryotes. 1996, Pubmed
Guadagno, Cdk2 kinase is required for entry into mitosis as a positive regulator of Cdc2-cyclin B kinase activity. 1996, Pubmed , Xenbase
Heichman, The yeast CDC16 and CDC27 genes restrict DNA replication to once per cell cycle. 1996, Pubmed
Hendrickson, Phosphorylation of MCM4 by cdc2 protein kinase inhibits the activity of the minichromosome maintenance complex. 1996, Pubmed , Xenbase
Hennessy, A group of interacting yeast DNA replication genes. 1991, Pubmed
Hereford, Sequential gene function in the initiation of Saccharomyces cerevisiae DNA synthesis. 1975, Pubmed
Hua, A role for Cdk2 kinase in negatively regulating DNA replication during S phase of the cell cycle. 1997, Pubmed , Xenbase
Jallepalli, Rum1 and Cdc18 link inhibition of cyclin-dependent kinase to the initiation of DNA replication in Schizosaccharomyces pombe. 1996, Pubmed
Kelly, The fission yeast cdc18+ gene product couples S phase to START and mitosis. 1993, Pubmed
Kominami, Fission yeast WD-repeat protein pop1 regulates genome ploidy through ubiquitin-proteasome-mediated degradation of the CDK inhibitor Rum1 and the S-phase initiator Cdc18. 1997, Pubmed
Kornbluth, In vitro cell cycle arrest induced by using artificial DNA templates. 1992, Pubmed , Xenbase
Kubota, Identification of the yeast MCM3-related protein as a component of Xenopus DNA replication licensing factor. 1995, Pubmed , Xenbase
Li, Isolation of ORC6, a component of the yeast origin recognition complex by a one-hybrid system. 1994, Pubmed
Liang, ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome. 1995, Pubmed
Madine, MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells. 1995, Pubmed , Xenbase
Madine, The nuclear envelope prevents reinitiation of replication by regulating the binding of MCM3 to chromatin in Xenopus egg extracts. 1996, Pubmed , Xenbase
Muzi Falconi, cdc18+ regulates initiation of DNA replication in Schizosaccharomyces pombe. 1996, Pubmed
Newport, Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. 1987, Pubmed , Xenbase
Nishitani, p65cdc18 plays a major role controlling the initiation of DNA replication in fission yeast. 1996, Pubmed
Piatti, Cdc6 is an unstable protein whose de novo synthesis in G1 is important for the onset of S phase and for preventing a 'reductional' anaphase in the budding yeast Saccharomyces cerevisiae. 1995, Pubmed
Piatti, Activation of S-phase-promoting CDKs in late G1 defines a "point of no return" after which Cdc6 synthesis cannot promote DNA replication in yeast. 1996, Pubmed
Rao, The origin recognition complex interacts with a bipartite DNA binding site within yeast replicators. 1995, Pubmed
Rowles, Interaction between the origin recognition complex and the replication licensing system in Xenopus. 1996, Pubmed , Xenbase
Roy, Activation of p34cdc2 kinase by cyclin A. 1991, Pubmed , Xenbase
Santocanale, ORC- and Cdc6-dependent complexes at active and inactive chromosomal replication origins in Saccharomyces cerevisiae. 1997, Pubmed
Schulte, Expression, phosphorylation and nuclear localization of the human P1 protein, a homologue of the yeast Mcm 3 replication protein. 1995, Pubmed
Sheehan, Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs. 1988, Pubmed , Xenbase
Strausfeld, Both cyclin A and cyclin E have S-phase promoting (SPF) activity in Xenopus egg extracts. 1997, Pubmed , Xenbase
Strausfeld, Cip1 blocks the initiation of DNA replication in Xenopus extracts by inhibition of cyclin-dependent kinases. 1995, Pubmed , Xenbase
Tanaka, Loading of an Mcm protein onto DNA replication origins is regulated by Cdc6p and CDKs. 1997, Pubmed
Todorov, BM28, a human member of the MCM2-3-5 family, is displaced from chromatin during DNA replication. 1995, Pubmed
Walter, Regulation of replicon size in Xenopus egg extracts. 1997, Pubmed , Xenbase
Wuarin, Regulating S phase: CDKs, licensing and proteolysis. 1996, Pubmed
Yan, Cell cycle-regulated nuclear localization of MCM2 and MCM3, which are required for the initiation of DNA synthesis at chromosomal replication origins in yeast. 1993, Pubmed
Yan, Mcm2 and Mcm3, two proteins important for ARS activity, are related in structure and function. 1991, Pubmed
Yew, Proteolysis and DNA replication: the CDC34 requirement in the Xenopus egg cell cycle. 1997, Pubmed , Xenbase
Yoon, Regulation of Saccharomyces cerevisiae CDC7 function during the cell cycle. 1993, Pubmed
Zwerschke, The Saccharomyces cerevisiae CDC6 gene is transcribed at late mitosis and encodes a ATP/GTPase controlling S phase initiation. 1994, Pubmed