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J Biol Chem
2014 Dec 26;28952:35987-6000. doi: 10.1074/jbc.M114.589119.
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Protein phosphatase 2A and Cdc7 kinase regulate the DNA unwinding element-binding protein in replication initiation.
Gao Y
,
Yao J
,
Poudel S
,
Romer E
,
Abu-Niaaj L
,
Leffak M
.
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The DNA unwinding element (DUE)-binding protein (DUE-B) binds to replication origins coordinately with the minichromosome maintenance (MCM) helicase and the helicase activator Cdc45 in vivo, and loads Cdc45 onto chromatin in Xenopus egg extracts. Human DUE-B also retains the aminoacyl-tRNA proofreading function of its shorter orthologs in lower organisms. Here we report that phosphorylation of the DUE-B unstructured C-terminal domain unique to higher organisms regulates DUE-B intermolecular binding. Gel filtration analyses show that unphosphorylated DUE-B forms multiple high molecular weight (HMW) complexes. Several aminoacyl-tRNA synthetases and Mcm2-7 proteins were identified by mass spectrometry of the HMW complexes. Aminoacyl-tRNA synthetase binding is RNase A sensitive, whereas interaction with Mcm2-7 is nuclease resistant. Unphosphorylated DUE-B HMW complex formation is decreased by PP2A inhibition or direct DUE-B phosphorylation, and increased by inhibition of Cdc7. These results indicate that the state of DUE-B phosphorylation is maintained by the equilibrium between Cdc7-dependent phosphorylation and PP2A-dependent dephosphorylation, each previously shown to regulate replication initiation. Alanine mutation of the DUE-B C-terminal phosphorylation target sites increases MCM binding but blocks Cdc45 loading in vivo and inhibits cell division. In egg extracts alanine mutation of the DUE-B C-terminal phosphorylation sites blocks Cdc45 loading and inhibits DNA replication. The effects of DUE-B C-terminal phosphorylation reveal a novel S phase kinase regulatory mechanism for Cdc45 loading and MCM helicase activation.
Aladjem,
Genetic dissection of a mammalian replicator in the human beta-globin locus.
1998, Pubmed
Aladjem,
Genetic dissection of a mammalian replicator in the human beta-globin locus.
1998,
Pubmed
Balestrini,
GEMC1 is a TopBP1-interacting protein required for chromosomal DNA replication.
2010,
Pubmed
,
Xenbase
Bell,
ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex.
1992,
Pubmed
Bialojan,
Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Specificity and kinetics.
1988,
Pubmed
Casper,
The c-myc DNA-unwinding element-binding protein modulates the assembly of DNA replication complexes in vitro.
2005,
Pubmed
,
Xenbase
Charych,
Inhibition of Cdc7/Dbf4 kinase activity affects specific phosphorylation sites on MCM2 in cancer cells.
2008,
Pubmed
Chen,
Activation of a human chromosomal replication origin by protein tethering.
2013,
Pubmed
Cho,
CDC7 kinase phosphorylates serine residues adjacent to acidic amino acids in the minichromosome maintenance 2 protein.
2006,
Pubmed
Chong,
Characterization of the Xenopus replication licensing system.
1997,
Pubmed
,
Xenbase
Chou,
Protein phosphatase 2A regulates binding of Cdc45 to the prereplication complex.
2002,
Pubmed
,
Xenbase
Chowdhury,
The DNA unwinding element binding protein DUE-B interacts with Cdc45 in preinitiation complex formation.
2010,
Pubmed
,
Xenbase
Cohen,
An improved procedure for identifying and quantitating protein phosphatases in mammalian tissues.
1989,
Pubmed
Coudreuse,
Driving the cell cycle with a minimal CDK control network.
2010,
Pubmed
Evrin,
In the absence of ATPase activity, pre-RC formation is blocked prior to MCM2-7 hexamer dimerization.
2013,
Pubmed
Fernández-Cid,
An ORC/Cdc6/MCM2-7 complex is formed in a multistep reaction to serve as a platform for MCM double-hexamer assembly.
2013,
Pubmed
Gambus,
MCM2-7 form double hexamers at licensed origins in Xenopus egg extract.
2011,
Pubmed
,
Xenbase
Ghosh,
Differential binding of replication proteins across the human c-myc replicator.
2006,
Pubmed
Hardy,
mcm5/cdc46-bob1 bypasses the requirement for the S phase activator Cdc7p.
1997,
Pubmed
Hausmann,
Aminoacyl-tRNA synthetase complexes: molecular multitasking revealed.
2008,
Pubmed
Heller,
Eukaryotic origin-dependent DNA replication in vitro reveals sequential action of DDK and S-CDK kinases.
2011,
Pubmed
Hoang,
Structural changes in Mcm5 protein bypass Cdc7-Dbf4 function and reduce replication origin efficiency in Saccharomyces cerevisiae.
2007,
Pubmed
Ishimi,
Enhanced expression of Mcm proteins in cancer cells derived from uterine cervix.
2003,
Pubmed
Kamimura,
Sld3, which interacts with Cdc45 (Sld4), functions for chromosomal DNA replication in Saccharomyces cerevisiae.
2001,
Pubmed
Kemp,
Structure and function of the c-myc DNA-unwinding element-binding protein DUE-B.
2007,
Pubmed
Kim,
Protein-protein interactions and multi-component complexes of aminoacyl-tRNA synthetases.
2014,
Pubmed
Krasinska,
Protein phosphatase 2A controls the order and dynamics of cell-cycle transitions.
2011,
Pubmed
,
Xenbase
Kumagai,
Treslin collaborates with TopBP1 in triggering the initiation of DNA replication.
2010,
Pubmed
,
Xenbase
Kumagai,
Direct regulation of Treslin by cyclin-dependent kinase is essential for the onset of DNA replication.
2011,
Pubmed
,
Xenbase
Labib,
How do Cdc7 and cyclin-dependent kinases trigger the initiation of chromosome replication in eukaryotic cells?
2010,
Pubmed
Lebofsky,
DNA replication in nucleus-free Xenopus egg extracts.
2009,
Pubmed
,
Xenbase
Lee,
Aminoacyl-tRNA synthetase complexes: beyond translation.
2004,
Pubmed
Lin,
Protein phosphatase 2A is required for the initiation of chromosomal DNA replication.
1998,
Pubmed
,
Xenbase
Liu,
Unstable spinocerebellar ataxia type 10 (ATTCT*(AGAAT) repeats are associated with aberrant replication at the ATX10 locus and replication origin-dependent expansion at an ectopic site in human cells.
2007,
Pubmed
Masai,
Phosphorylation of MCM4 by Cdc7 kinase facilitates its interaction with Cdc45 on the chromatin.
2006,
Pubmed
Montagnoli,
Identification of Mcm2 phosphorylation sites by S-phase-regulating kinases.
2006,
Pubmed
Montagnoli,
A Cdc7 kinase inhibitor restricts initiation of DNA replication and has antitumor activity.
2008,
Pubmed
Muramatsu,
CDK-dependent complex formation between replication proteins Dpb11, Sld2, Pol (epsilon}, and GINS in budding yeast.
2010,
Pubmed
Petersen,
Protein phosphatase 2A antagonizes ATM and ATR in a Cdk2- and Cdc7-independent DNA damage checkpoint.
2006,
Pubmed
,
Xenbase
Randell,
Mec1 is one of multiple kinases that prime the Mcm2-7 helicase for phosphorylation by Cdc7.
2010,
Pubmed
Ruzzene,
Protein kinase CK2 inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) induces apoptosis and caspase-dependent degradation of haematopoietic lineage cell-specific protein 1 (HS1) in Jurkat cells.
2002,
Pubmed
Sansam,
A vertebrate gene, ticrr, is an essential checkpoint and replication regulator.
2010,
Pubmed
Sheu,
Cdc7-Dbf4 phosphorylates MCM proteins via a docking site-mediated mechanism to promote S phase progression.
2006,
Pubmed
Sheu,
The Dbf4-Cdc7 kinase promotes S phase by alleviating an inhibitory activity in Mcm4.
2010,
Pubmed
Walter,
Evidence for sequential action of cdc7 and cdk2 protein kinases during initiation of DNA replication in Xenopus egg extracts.
2000,
Pubmed
,
Xenbase
Weinreich,
Cdc7p-Dbf4p kinase binds to chromatin during S phase and is regulated by both the APC and the RAD53 checkpoint pathway.
1999,
Pubmed
Zegerman,
Phosphorylation of Sld2 and Sld3 by cyclin-dependent kinases promotes DNA replication in budding yeast.
2007,
Pubmed