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Switch on the engine: how the eukaryotic replicative helicase MCM2–7 becomes activated

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Abstract

A crucial step during eukaryotic initiation of DNA replication is the correct loading and activation of the replicative DNA helicase, which ensures that each replication origin fires only once. Unregulated DNA helicase loading and activation, as it occurs in cancer, can cause severe DNA damage and genomic instability. The essential mini-chromosome maintenance proteins 2–7 (MCM2–7) represent the core of the eukaryotic replicative helicase that is loaded at DNA replication origins during G1-phase of the cell cycle. The MCM2–7 helicase activity, however, is only triggered during S-phase once the holo-helicase Cdc45-MCM2–7-GINS (CMG) has been formed. A large number of factors and several kinases interact and contribute to CMG formation and helicase activation, though the exact mechanisms remain unclear. Crucially, upon DNA damage, this reaction is temporarily halted to ensure genome integrity. Here, we review the current understanding of helicase activation; we focus on protein interactions during CMG formation, discuss structural changes during helicase activation, and outline similarities and differences of the prokaryotic and eukaryotic helicase activation process.

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References

  • Ang XL, Wade Harper J (2005) SCF-mediated protein degradation and cell cycle control. Oncogene 24:2860–2870. doi:10.1038/sj.onc.1208614

    CAS  PubMed  Google Scholar 

  • Arias-Palomo E, O'Shea VL, Hood IV, Berger JM (2013) The bacterial DnaC helicase loader is a DnaB ring breaker. Cell 153:438–448. doi:10.1016/j.cell.2013.03.006

    PubMed Central  CAS  PubMed  Google Scholar 

  • Aucher W et al (2010) A strategy for interaction site prediction between phospho-binding modules and their partners identified from proteomic data. Mol Cell Proteomics 9:2745–2759. doi:10.1074/mcp.M110.003319

  • Balestrini A, Cosentino C, Errico A, Garner E, Costanzo V (2010) GEMC1 is a TopBP1-interacting protein required for chromosomal DNA replication. Nat Cell Biol 12:484–491

    PubMed Central  CAS  PubMed  Google Scholar 

  • Bell SP, Stillman B (1992) ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature 357:128–134. doi:10.1038/357128a0

    CAS  PubMed  Google Scholar 

  • Bochman ML, Schwacha A (2008) The Mcm2-7 complex has in vitro helicase activity. Mol Cell 31:287–293

    CAS  PubMed  Google Scholar 

  • Boos D, Sanchez-Pulido L, Rappas M, Pearl LH, Oliver AW, Ponting CP, Diffley JF (2011) Regulation of DNA replication through Sld3-Dpb11 interaction is conserved from yeast to humans. Curr Biol 21:1152–1157

    CAS  PubMed  Google Scholar 

  • Boos D, Frigola J, Diffley JF (2012) Activation of the replicative DNA helicase: breaking up is hard to do. Curr Opin Cell Biol 24:423–430. doi:10.1016/j.ceb.2012.01.011

    CAS  PubMed  Google Scholar 

  • Boos D, Yekezare M, Diffley JF (2013) Identification of a heteromeric complex that promotes DNA replication origin firing in human cells. Science 340:981–984. doi:10.1126/science.1237448

    CAS  PubMed  Google Scholar 

  • Bruck I, Kanter DM, Kaplan DL (2011) Enabling association of the GINS protein tetramer with the mini chromosome maintenance (Mcm)2-7 protein complex by phosphorylated Sld2 protein and single-stranded origin DNA. J Biol Chem 286:36414–36426

  • Bruck I, Kaplan D (2009) Dbf4-Cdc7 phosphorylation of Mcm2 is required for cell growth. J Biol Chem 284:28823–28831

  • Bruck I, Kaplan DL (2011a) GINS and Sld3 compete with one another for Mcm2-7 and Cdc45 binding. J Biol Chem 286:14157–14167

  • Bruck I, Kaplan DL (2011b) Origin single-stranded DNA releases Sld3 protein from the Mcm2-7 complex, allowing the GINS tetramer to bind the Mcm2-7 complex. J Biol Chem 286:18602–18613

  • Bruck I, Kaplan DL (2013) Cdc45 protein-single-stranded DNA interaction is important for stalling the helicase during replication stress. J Biol Chem 288:7550–7563

    PubMed Central  CAS  PubMed  Google Scholar 

  • Brummer A, Salazar C, Zinzalla V, Alberghina L, Hofer T (2010) Mathematical modelling of DNA replication reveals a trade-off between coherence of origin activation and robustness against rereplication. PLoS Comput Biol 6:e1000783. doi:10.1371/journal.pcbi.1000783

    PubMed Central  PubMed  Google Scholar 

  • Charych DH et al (2008) Inhibition of Cdc7/Dbf4 kinase activity affects specific phosphorylation sites on MCM2 in cancer cells. J Cell Biochem 104:1075–1086

    CAS  PubMed  Google Scholar 

  • Chen S, Bell SP (2011) CDK prevents Mcm2-7 helicase loading by inhibiting Cdt1 interaction with Orc6. Genes Dev 25:363–372. doi:10.1101/gad.2011511

    PubMed Central  CAS  PubMed  Google Scholar 

  • Chen S, de Vries MA, Bell SP (2007) Orc6 is required for dynamic recruitment of Cdt1 during repeated Mcm2-7 loading. Genes Dev 21:2897–2907

  • Cho WH, Lee YJ, Kong SI, Hurwitz J, Lee JK (2006) CDC7 kinase phosphorylates serine residues adjacent to acidic amino acids in the minichromosome maintenance 2 protein. Proc Natl Acad Sci U S A 103:11521–11526

    PubMed Central  CAS  PubMed  Google Scholar 

  • Chowdhury A et al (2010) The DNA unwinding element binding protein DUE-B interacts with Cdc45 in preinitiation complex formation. Mol Cell Biol 30:1495–1507

    PubMed Central  CAS  PubMed  Google Scholar 

  • Coleman TR, Carpenter PB, Dunphy WG (1996) The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts. Cell 87:53–63

    CAS  PubMed  Google Scholar 

  • Costa A, Ilves I, Tamberg N, Petojevic T, Nogales E, Botchan MR, Berger JM (2011) The structural basis for MCM2-7 helicase activation by GINS and Cdc45. Nat Struct Mol Biol 18:471–477. doi:10.1038/nsmb.2004

    PubMed Central  CAS  PubMed  Google Scholar 

  • Costa A et al (2014) DNA binding polarity, dimerization, and ATPase ring remodeling in the CMG helicase of the eukaryotic replisome. Elife 3:e03273. doi:10.7554/eLife.03273

    PubMed  Google Scholar 

  • Dave A, Cooley C, Garg M, Bianchi A (2014) Protein phosphatase 1 recruitment by Rif1 regulates DNA replication origin firing by counteracting DDK activity. Cell Rep 7:53–61. doi:10.1016/j.celrep.2014.02.019

    PubMed Central  CAS  PubMed  Google Scholar 

  • Di Perna R, Aria V, De Falco M, Sannino V, Okorokov AL, Pisani FM, De Felice M (2013) The physical interaction of Mcm10 with Cdc45 modulates their DNA-binding properties. Biochem J 454:333–343

    PubMed  Google Scholar 

  • Diffley JF (2010) The many faces of redundancy in DNA replication control. Cold Spring Harb Symp Quant Biol 75:135–142. doi:10.1101/sqb.2010.75.062

    CAS  PubMed  Google Scholar 

  • Donovan S, Harwood J, Drury LS, Diffley JF (1997) Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast. Proc Natl Acad Sci U S A 94:5611–5616

    PubMed Central  CAS  PubMed  Google Scholar 

  • Drury LS, Perkins G, Diffley JF (1997) The Cdc4/34/53 pathway targets Cdc6p for proteolysis in budding yeast. Embo J 16:5966–5976

    PubMed Central  CAS  PubMed  Google Scholar 

  • Duch A, Palou G, Jonsson ZO, Palou R, Calvo E, Wohlschlegel J, Quintana DG (2011) A Dbf4 mutant contributes to bypassing the Rad53-mediated block of origins of replication in response to genotoxic stress. J Biol Chem 286:2486–2491

    PubMed Central  CAS  PubMed  Google Scholar 

  • Edwards AM, Kus B, Jansen R, Greenbaum D, Greenblatt J, Gerstein M (2002) Bridging structural biology and genomics: assessing protein interaction data with known complexes. Trends Genet 18:529–536

  • Elsasser S, Chi Y, Yang P, Campbell JL (1999) Phosphorylation controls timing of Cdc6p destruction: a biochemical analysis. Mol Biol Cell 10:3263–3277

    PubMed Central  CAS  PubMed  Google Scholar 

  • Evrin C et al (2009) A double-hexameric MCM2-7 complex is loaded onto origin DNA during licensing of eukaryotic DNA replication. Proc Natl Acad Sci U S A 106:20240–20245

    PubMed Central  CAS  PubMed  Google Scholar 

  • Evrin C et al (2013) In the absence of ATPase activity, pre-RC formation is blocked prior to MCM2-7 hexamer dimerization. Nucleic Acids Res 41:3162–3172

    PubMed Central  CAS  PubMed  Google Scholar 

  • Evrin C et al (2014) The ORC/Cdc6/MCM2-7 complex facilitates MCM2-7 dimerization during prereplicative complex formation. Nucleic Acids Res 42:2257–2269

    PubMed Central  CAS  PubMed  Google Scholar 

  • Fernandez-Cid A et al (2013) An ORC/Cdc6/MCM2-7 complex is formed in a multistep reaction to serve as a platform for MCM double-hexamer assembly. Mol Cell 50:577–588

    CAS  PubMed  Google Scholar 

  • Ferreira MF, Santocanale C, Drury LS, Diffley JF (2000) Dbf4p, an essential S phase-promoting factor, is targeted for degradation by the anaphase-promoting complex. Mol Cell Biol 20:242–248

    CAS  PubMed  Google Scholar 

  • Francis LI, Randell JC, Takara TJ, Uchima L, Bell SP (2009) Incorporation into the prereplicative complex activates the Mcm2-7 helicase for Cdc7-Dbf4 phosphorylation. Genes Dev 23:643–654

    PubMed Central  CAS  PubMed  Google Scholar 

  • Frigola J, Remus D, Mehanna A, Diffley JF (2013) ATPase-dependent quality control of DNA replication origin licensing. Nature 495:339–343. doi:10.1038/nature11920

    CAS  PubMed  Google Scholar 

  • Froelich CA, Kang S, Epling LB, Bell SP, Enemark EJ (2014) A conserved MCM single-stranded DNA binding element is essential for replication initiation. Elife 3:e01993. doi:10.7554/eLife.01993

    PubMed Central  PubMed  Google Scholar 

  • Fu YV et al (2011) Selective bypass of a lagging strand roadblock by the eukaryotic replicative DNA helicase. Cell 146:931–941. doi:10.1016/j.cell.2011.07.045

    PubMed Central  CAS  PubMed  Google Scholar 

  • Fukuura M, Nagao K, Obuse C, Takahashi TS, Nakagawa T, Masukata H (2011) CDK promotes interactions of Sld3 and Drc1 with Cut5 for initiation of DNA replication in fission yeast. Mol Biol Cell 22:2620–2633

    PubMed Central  CAS  PubMed  Google Scholar 

  • Gambus A, Jones RC, Sanchez-Diaz A, Kanemaki M, van Deursen F, Edmondson RD, Labib K (2006) GINS maintains association of Cdc45 with MCM in replisome progression complexes at eukaryotic DNA replication forks. Nat Cell Biol 8:358–366

    CAS  PubMed  Google Scholar 

  • Gambus A et al (2009) A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome. Embo J 28:2992–3004

    PubMed Central  CAS  PubMed  Google Scholar 

  • Gambus A, Khoudoli GA, Jones RC, Blow JJ (2011) MCM2-7 form double hexamers at licensed origins in Xenopus egg extract. J Biol Chem 286:11855–11864

    PubMed Central  CAS  PubMed  Google Scholar 

  • Garcia V, Furuya K, Carr AM (2005) Identification and functional analysis of TopBP1 and its homologs. DNA Repair (Amst) 4:1227–1239

    CAS  Google Scholar 

  • Hardy CF, Dryga O, Seematter S, Pahl PM, Sclafani RA (1997) mcm5/cdc46-bob1 bypasses the requirement for the S phase activator Cdc7p. Proc Natl Acad Sci U S A 94:3151–3155

    PubMed Central  CAS  PubMed  Google Scholar 

  • Hashimoto Y, Takisawa H (2003) Xenopus Cut5 is essential for a CDK-dependent process in the initiation of DNA replication. Embo J 22:2526–2535

    PubMed Central  CAS  PubMed  Google Scholar 

  • Hashimoto Y, Puddu F, Costanzo V (2012) RAD51- and MRE11-dependent reassembly of uncoupled CMG helicase complex at collapsed replication forks. Nat Struct Mol Biol 19:17–24

    PubMed Central  CAS  Google Scholar 

  • Hayano M, Kanoh Y, Matsumoto S, Renard-Guillet C, Shirahige K, Masai H (2012) Rif1 is a global regulator of timing of replication origin firing in fission yeast. Genes Dev 26:137–150

    PubMed Central  CAS  PubMed  Google Scholar 

  • Heller RC, Kang S, Lam WM, Chen S, Chan CS, Bell SP (2011) Eukaryotic origin-dependent DNA replication in vitro reveals sequential action of DDK and S-CDK kinases. Cell 146:80–91. doi:10.1016/j.cell.2011.06.012

    PubMed Central  CAS  PubMed  Google Scholar 

  • Hiraga S et al (2014) Rif1 controls DNA replication by directing protein phosphatase 1 to reverse Cdc7-mediated phosphorylation of the MCM complex. Genes Dev 28:372–383

    PubMed Central  CAS  PubMed  Google Scholar 

  • Humphrey T, Pearce A (2005) Cell cycle molecules and mechanisms of the budding and fission yeasts. Methods Mol Biol 296:3–29

    CAS  PubMed  Google Scholar 

  • Huo L et al (2012) The Rix1 (Ipi1p-2p-3p) complex is a critical determinant of DNA replication licensing independent of their roles in ribosome biogenesis. Cell Cycle 11:1325–1339

    CAS  PubMed  Google Scholar 

  • Ilves I, Petojevic T, Pesavento JJ, Botchan MR (2010) Activation of the MCM2-7 helicase by association with Cdc45 and GINS proteins. Mol Cell 37:247–258

    CAS  PubMed  Google Scholar 

  • Im JS, Ki SH, Farina A, Jung DS, Hurwitz J, Lee JK (2009) Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins. Proc Natl Acad Sci U S A 106:15628–15632

    PubMed Central  CAS  PubMed  Google Scholar 

  • Ishimi Y (1997) A DNA helicase activity is associated with an MCM4, −6, and −7 protein complex. J Biol Chem 272:24508–24513

    CAS  PubMed  Google Scholar 

  • Itou H, Muramatsu S, Shirakihara Y, Araki H (2014) Crystal structure of the homology domain of the eukaryotic DNA replication proteins sld3/treslin. Structure 22:1341–1347

    CAS  PubMed  Google Scholar 

  • Jaspersen SL, Charles JF, Morgan DO (1999) Inhibitory phosphorylation of the APC regulator Hct1 is controlled by the kinase Cdc28 and the phosphatase Cdc14. Curr Biol 9:227–236

    CAS  PubMed  Google Scholar 

  • Kamimura Y, Masumoto H, Sugino A, Araki H (1998) Sld2, which interacts with Dpb11 in Saccharomyces cerevisiae, is required for chromosomal DNA replication. Mol Cell Biol 18:6102–6109

  • Kamimura Y, Tak YS, Sugino A, Araki H (2001) Sld3, which interacts with Cdc45 (Sld4), functions for chromosomal DNA replication in Saccharomyces cerevisiae. Embo J 20:2097–2107

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kanemaki M, Labib K (2006) Distinct roles for Sld3 and GINS during establishment and progression of eukaryotic DNA replication forks. Embo J 25:1753–1763

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kang YH, Farina A, Bermudez VP, Tappin I, Du F, Galal WC, Hurwitz J (2013) Interaction between human Ctf4 and the Cdc45/Mcm2-7/GINS (CMG) replicative helicase. Proc Natl Acad Sci U S A 110:19760–19765

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kanke M, Kodama Y, Takahashi TS, Nakagawa T, Masukata H (2012) Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components. Embo J 31:2182–2194

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kanter DM, Kaplan DL (2011) Sld2 binds to origin single-stranded DNA and stimulates DNA annealing. Nucleic Acids Res 39:2580–2592

    PubMed Central  CAS  PubMed  Google Scholar 

  • Katou Y, Kaneshiro K, Aburatani H, Shirahige K (2006) Genomic approach for the understanding of dynamic aspect of chromosome behavior. Methods Enzymol 409:389–410. doi:10.1016/S0076-6879(05)09023-3

    CAS  PubMed  Google Scholar 

  • Krastanova I, Sannino V, Amenitsch H, Gileadi O, Pisani FM, Onesti S (2012) Structural and functional insights into the DNA replication factor Cdc45 reveal an evolutionary relationship to the DHH family of phosphoesterases. J Biol Chem 287:4121–4128

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kumagai A, Shevchenko A, Dunphy WG (2010) Treslin collaborates with TopBP1 in triggering the initiation of DNA replication. Cell 140:349–359

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kumagai A, Shevchenko A, Dunphy WG (2011) Direct regulation of Treslin by cyclin-dependent kinase is essential for the onset of DNA replication. J Cell Biol 193:995–1007. doi:10.1083/jcb.201102003

    PubMed Central  CAS  PubMed  Google Scholar 

  • Kunkel TA, Burgers PM (2008) Dividing the workload at a eukaryotic replication fork Trends. Cell Biol 18:521–527. doi:10.1016/j.tcb.2008.08.005

    CAS  Google Scholar 

  • Labib K (2010) How do Cdc7 and cyclin-dependent kinases trigger the initiation of chromosome replication in eukaryotic cells? Genes Dev 24:1208–1219. doi:10.1101/gad.1933010

    PubMed Central  CAS  PubMed  Google Scholar 

  • Labib K, Diffley JF, Kearsey SE (1999) G1-phase and B-type cyclins exclude the DNA-replication factor Mcm4 from the nucleus. Nat Cell Biol 1:415–422

    CAS  PubMed  Google Scholar 

  • Laskey RA, Madine MA (2003) A rotary pumping model for helicase function of MCM proteins at a distance from replication forks. EMBO Rep 4:26–30. doi:10.1038/sj.embor.embor706

    PubMed Central  CAS  PubMed  Google Scholar 

  • Lee C, Liachko I, Bouten R, Kelman Z, Tye BK (2010) Alternative mechanisms for coordinating polymerase alpha and MCM helicase. Mol Cell Biol 30:423–435. doi:10.1128/MCB.01240-09

  • Lee JK, Hurwitz J (2000) Isolation and characterization of various complexes of the minichromosome maintenance proteins of Schizosaccharomyces pombe. J Biol Chem 275:18871–18878. doi:10.1074/jbc.M001118200

    CAS  PubMed  Google Scholar 

  • Liachko I, Tye BK (2009) Mcm10 mediates the interaction between DNA replication and silencing machineries. Genetics 181:379–391

  • Liang C, Stillman B (1997) Persistent initiation of DNA replication and chromatin-bound MCM proteins during the cell cycle in cdc6 mutants. Genes Dev 11:3375–3386

    PubMed Central  CAS  PubMed  Google Scholar 

  • Liku ME, Nguyen VQ, Rosales AW, Irie K, Li JJ (2005) CDK phosphorylation of a novel NLS-NES module distributed between two subunits of the Mcm2-7 complex prevents chromosomal rereplication. Mol Biol Cell 16:5026–5039

    PubMed Central  CAS  PubMed  Google Scholar 

  • Lopez-Mosqueda J, Maas NL, Jonsson ZO, Defazio-Eli LG, Wohlschlegel J, Toczyski DP (2010) Damage-induced phosphorylation of Sld3 is important to block late origin firing. Nature 467:479–483

    PubMed Central  CAS  PubMed  Google Scholar 

  • Makiniemi M et al (2001) BRCT domain-containing protein TopBP1 functions in DNA replication and damage response. J Biol Chem 276:30399–30406

    CAS  PubMed  Google Scholar 

  • Masai H, Matsui E, You Z, Ishimi Y, Tamai K, Arai K (2000) Human Cdc7-related kinase complex. In vitro phosphorylation of MCM by concerted actions of Cdks and Cdc7 and that of a criticial threonine residue of Cdc7 bY Cdks. J Biol Chem 275:29042–29052

    CAS  PubMed  Google Scholar 

  • Masai H et al (2006) Phosphorylation of MCM4 by Cdc7 kinase facilitates its interaction with Cdc45 on the chromatin. J Biol Chem 281:39249–39261

    CAS  PubMed  Google Scholar 

  • Masumoto H, Muramatsu S, Kamimura Y, Araki H (2002) S-Cdk-dependent phosphorylation of Sld2 essential for chromosomal DNA replication in budding yeast. Nature 415:651–655

    CAS  PubMed  Google Scholar 

  • Matsumoto S, Hayano M, Kanoh Y, Masai H (2011) Multiple pathways can bypass the essential role of fission yeast Hsk1 kinase in DNA replication initiation. J Cell Biol 195:387–401

    PubMed Central  CAS  PubMed  Google Scholar 

  • Matsuno K, Kumano M, Kubota Y, Hashimoto Y, Takisawa H (2006) The N-terminal noncatalytic region of Xenopus RecQ4 is required for chromatin binding of DNA polymerase alpha in the initiation of DNA replication. Mol Cell Biol 26:4843–4852

    PubMed Central  CAS  PubMed  Google Scholar 

  • Mattarocci S et al (2014) Rif1 controls DNA replication timing in yeast through the PP1 phosphatase Glc7. Cell Rep 7:62–69. doi:10.1016/j.celrep.2014.03.010

    CAS  PubMed  Google Scholar 

  • McIntosh D, Blow JJ (2012) Dormant origins, the licensing checkpoint, and the response to replicative stresses Cold Spring Harb Perspect Biol 4. doi:10.1101/cshperspect.a012955

  • Merchant AM, Kawasaki Y, Chen Y, Lei M, Tye BK (1997) A lesion in the DNA replication initiation factor Mcm10 induces pausing of elongation forks through chromosomal replication origins in Saccharomyces cerevisiae. Mol Cell Biol 17:3261–3271

  • Montagnoli A et al (2006) Identification of Mcm2 phosphorylation sites by S-phase-regulating kinases. J Biol Chem 281:10281–10290

    CAS  PubMed  Google Scholar 

  • Moyer SE, Lewis PW, Botchan MR (2006) Isolation of the Cdc45/Mcm2-7/GINS (CMG) complex, a candidate for the eukaryotic DNA replication fork helicase. Proc Natl Acad Sci U S A 103:10236–10241

    PubMed Central  CAS  PubMed  Google Scholar 

  • Muramatsu S, Hirai K, Tak YS, Kamimura Y, Araki H (2010) CDK-dependent complex formation between replication proteins Dpb11, Sld2, Pol (epsilon}, and GINS in budding yeast. Genes Dev 24:602–612

    PubMed Central  CAS  PubMed  Google Scholar 

  • Nakajima R, Masukata H (2002) SpSld3 is required for loading and maintenance of SpCdc45 on chromatin in DNA replication in fission yeast. Mol Biol Cell 13:1462–1472

    PubMed Central  CAS  PubMed  Google Scholar 

  • Natsume T et al (2013) Kinetochores coordinate pericentromeric cohesion and early DNA replication by Cdc7-Dbf4 kinase recruitment. Mol Cell 50:661–674

    PubMed Central  CAS  PubMed  Google Scholar 

  • Nguyen VQ, Co C, Li JJ (2001) Cyclin-dependent kinases prevent DNA re-replication through multiple mechanisms. Nature 411:1068–1073. doi:10.1038/35082600

    CAS  PubMed  Google Scholar 

  • Noguchi E, Shanahan P, Noguchi C, Russell P (2002) CDK phosphorylation of Drc1 regulates DNA replication in fission yeast. Curr Biol 12:599–605

    CAS  PubMed  Google Scholar 

  • On KF, Beuron F, Frith D, Snijders AP, Morris EP, Diffley JF (2014) Prereplicative complexes assembled in vitro support origin-dependent and independent DNA replication. Embo J 33:605–620. doi:10.1002/embj.201387369

    PubMed Central  CAS  PubMed  Google Scholar 

  • Oshiro G, Owens JC, Shellman Y, Sclafani RA, Li JJ (1999) Cell cycle control of Cdc7p kinase activity through regulation of Dbf4p stability. Mol Cell Biol 19:4888–4896

    PubMed Central  CAS  PubMed  Google Scholar 

  • Perkins G, Drury LS, Diffley JF (2001) Separate SCF(CDC4) recognition elements target Cdc6 for proteolysis in S phase and mitosis. Embo J 20:4836–4845

    PubMed Central  CAS  PubMed  Google Scholar 

  • Peters JM (2006) The anaphase promoting complex/cyclosome: a machine designed to destroy. Nat Rev Mol Cell Biol 7:644–656. doi:10.1038/nrm1988

    CAS  PubMed  Google Scholar 

  • Ramer MD, Suman ES, Richter H, Stanger K, Spranger M, Bieberstein N, Duncker BP (2013) Dbf4 and Cdc7 proteins promote DNA replication through interactions with distinct Mcm2-7 protein subunits. J Biol Chem 288:14926–14935

    PubMed Central  CAS  PubMed  Google Scholar 

  • Randell JC, Bowers JL, Rodriguez HK, Bell SP (2006) Sequential ATP hydrolysis by Cdc6 and ORC directs loading of the Mcm2-7 helicase. Mol Cell 21:29–39

    CAS  PubMed  Google Scholar 

  • Randell JC, Fan A, Chan C, Francis LI, Heller RC, Galani K, Bell SP (2010) Mec1 is one of multiple kinases that prime the Mcm2-7 helicase for phosphorylation by Cdc7. Mol Cell 40:353–363. doi:10.1016/j.molcel.2010.10.017

    PubMed Central  CAS  PubMed  Google Scholar 

  • Rappas M, Oliver AW, Pearl LH (2011) Structure and function of the Rad9-binding region of the DNA-damage checkpoint adaptor TopBP1. Nucleic Acids Res 39:313–324. doi:10.1093/nar/gkq743

    PubMed Central  CAS  PubMed  Google Scholar 

  • Remus D, Beuron F, Tolun G, Griffith JD, Morris EP, Diffley JF (2009) Concerted loading of Mcm2-7 double hexamers around DNA during DNA replication origin licensing. Cell 139:719–730

    PubMed Central  CAS  PubMed  Google Scholar 

  • Renard-Guillet C, Kanoh Y, Shirahige K, Masai H (2014) Temporal and spatial regulation of eukaryotic DNA replication: from regulated initiation to genome-scale timing program. Semin Cell Dev Biol 30C:110–120. doi:10.1016/j.semcdb.2014.04.014

    Google Scholar 

  • Riera A, Tognetti S, Speck C (2014) Helicase loading: how to build a MCM2-7 double-hexamer. Semin Cell Dev Biol 30C:104–109

    Google Scholar 

  • Saka Y, Fantes P, Sutani T, McInerny C, Creanor J, Yanagida M (1994) Fission yeast cut5 links nuclear chromatin and M phase regulator in the replication checkpoint control. Embo J 13:5319–5329

    PubMed Central  CAS  PubMed  Google Scholar 

  • Samel SA et al (2014) A unique DNA entry gate serves for regulated loading of the eukaryotic replicative helicase MCM2–7 onto DNA. Genes Dev 28:1653–1666

    PubMed Central  CAS  PubMed  Google Scholar 

  • Sanchez-Pulido L, Ponting CP (2011) Cdc45: the missing RecJ ortholog in eukaryotes? Bioinformatics 27:1885–1888. doi:10.1093/bioinformatics/btr332

    CAS  PubMed  Google Scholar 

  • Sangrithi MN, Bernal JA, Madine M, Philpott A, Lee J, Dunphy WG, Venkitaraman AR (2005) Initiation of DNA replication requires the RECQL4 protein mutated in Rothmund-Thomson syndrome. Cell 121:887–898

    CAS  PubMed  Google Scholar 

  • Schmidt U, Wollmann Y, Franke C, Grosse F, Saluz HP, Hanel F (2008) Characterization of the interaction between the human DNA topoisomerase IIbeta-binding protein 1 (TopBP1) and the cell division cycle 45 (Cdc45) protein. Biochem J 409:169–177

    CAS  PubMed  Google Scholar 

  • Sheu YJ, Stillman B (2006) Cdc7-Dbf4 phosphorylates MCM proteins via a docking site-mediated mechanism to promote S phase progression. Mol Cell 24:101–113

    PubMed Central  CAS  PubMed  Google Scholar 

  • Sheu YJ, Stillman B (2010) The Dbf4-Cdc7 kinase promotes S phase by alleviating an inhibitory activity in Mcm4. Nature 463:113–117. doi:10.1038/nature08647

    PubMed Central  CAS  PubMed  Google Scholar 

  • Siddiqui K, On KF, Diffley JF (2013) Regulating DNA replication in eukarya Cold Spring Harb Perspect Biol 5. doi:10.1101/cshperspect.a012930

  • Simon AC et al (2014) A Ctf4 trimer couples the CMG helicase to DNA polymerase alpha in the eukaryotic replisome. Nature. doi:10.1038/nature13234

    PubMed Central  Google Scholar 

  • Skarstad K, Katayama T (2013) Regulating DNA replication in bacteria. Cold Spring Harb Perspect Biol 5:a012922. doi:10.1101/cshperspect.a012922

    PubMed  Google Scholar 

  • Speck C, Messer W (2001) Mechanism of origin unwinding: sequential binding of DnaA to double- and single-stranded DNA. Embo J 20:1469–1476. doi:10.1093/emboj/20.6.1469

    PubMed Central  CAS  PubMed  Google Scholar 

  • Speck C, Chen Z, Li H, Stillman B (2005) ATPase-dependent cooperative binding of ORC and Cdc6 to origin DNA. Nat Struct Mol Biol 12:965–971

    PubMed Central  CAS  PubMed  Google Scholar 

  • Sun J et al (2013) Cryo-EM structure of a helicase loading intermediate containing ORC-Cdc6-Cdt1-MCM2-7 bound to DNA. Nat Struct Mol Biol 20:944–951

    PubMed Central  CAS  PubMed  Google Scholar 

  • Sun J et al (2014) Structural and mechanistic insights into Mcm2-7 double-hexamer assembly and function. Genes Dev 28: in press

  • Szambowska A, Tessmer I, Kursula P, Usskilat C, Prus P, Pospiech H, Grosse F (2014) DNA binding properties of human Cdc45 suggest a function as molecular wedge for DNA unwinding. Nucleic Acids Res 42:2308–2319

    PubMed Central  CAS  PubMed  Google Scholar 

  • Tak YS, Tanaka Y, Endo S, Kamimura Y, Araki H (2006) A CDK-catalysed regulatory phosphorylation for formation of the DNA replication complex Sld2-Dpb11. Embo J 25:1987–1996. doi:10.1038/sj.emboj.7601075

    PubMed Central  CAS  PubMed  Google Scholar 

  • Takara TJ, Bell SP (2011) Multiple Cdt1 molecules act at each origin to load replication-competent Mcm2-7 helicases. Embo J 30:4885–4896

  • Takayama Y, Kamimura Y, Okawa M, Muramatsu S, Sugino A, Araki H (2003) GINS, a novel multiprotein complex required for chromosomal DNA replication in budding yeast. Genes Dev 17:1153–1165

  • Tanaka H et al (2009) Ctf4 coordinates the progression of helicase and DNA polymerase alpha. Genes Cells 14:807–820

  • Tanaka S, Araki H (2013) Helicase activation and establishment of replication forks at chromosomal origins of replication. Cold Spring Harb Perspect Biol 5:a010371. doi:10.1101/cshperspect.a010371

    PubMed  Google Scholar 

  • Tanaka S, Diffley JF (2002) Interdependent nuclear accumulation of budding yeast Cdt1 and Mcm2-7 during G1 phase. Nat Cell Biol 4:198–207

    CAS  PubMed  Google Scholar 

  • Tanaka T, Knapp D, Nasmyth K (1997) Loading of an Mcm protein onto DNA replication origins is regulated by Cdc6p and CDKs. Cell 90:649–660

    CAS  PubMed  Google Scholar 

  • Tanaka S, Umemori T, Hirai K, Muramatsu S, Kamimura Y, Araki H (2007) CDK-dependent phosphorylation of Sld2 and Sld3 initiates DNA replication in budding yeast. Nature 445:328–332

    CAS  PubMed  Google Scholar 

  • Tanaka S, Nakato R, Katou Y, Shirahige K, Araki H (2011) Origin association of Sld3, Sld7, and Cdc45 proteins is a key step for determination of origin-firing timing. Curr Biol 21:2055–2063

    CAS  PubMed  Google Scholar 

  • Tanaka S, Komeda Y, Umemori T, Kubota Y, Takisawa H, Araki H (2013) Efficient initiation of DNA replication in eukaryotes requires Dpb11/TopBP1-GINS interaction. Mol Cell Biol 33:2614–2622

    PubMed Central  CAS  PubMed  Google Scholar 

  • van Deursen F, Sengupta S, De Piccoli G, Sanchez-Diaz A, Labib K (2012) Mcm10 associates with the loaded DNA helicase at replication origins and defines a novel step in its activation. Embo J 31:2195–2206

    PubMed Central  PubMed  Google Scholar 

  • Van Hatten RA, Tutter AV, Holway AH, Khederian AM, Walter JC, Michael WM (2002) The Xenopus Xmus101 protein is required for the recruitment of Cdc45 to origins of DNA replication. J Cell Biol 159:541–547

    PubMed Central  PubMed  Google Scholar 

  • Wang G, Tong X, Weng S, Zhou H (2012) Multiple phosphorylation of Rad9 by CDK is required for DNA damage checkpoint activation. Cell Cycle 11:3792–3800. doi:10.4161/cc.21987

  • Warren EM, Vaithiyalingam S, Haworth J, Greer B, Bielinsky AK, Chazin WJ, Eichman BF (2008) Structural basis for DNA binding by replication initiator Mcm10. Structure 16:1892–1901. doi:10.1016/j.str.2008.10.005

    PubMed Central  CAS  PubMed  Google Scholar 

  • Watase G, Takisawa H, Kanemaki MT (2012) Mcm10 plays a role in functioning of the eukaryotic replicative DNA helicase, Cdc45-Mcm-GINS. Curr Biol 22:343–349

    CAS  PubMed  Google Scholar 

  • Weinreich M, Stillman B (1999) Cdc7p-Dbf4p kinase binds to chromatin during S phase and is regulated by both the APC and the RAD53 checkpoint pathway. Embo J 18:5334–5346

    PubMed Central  CAS  PubMed  Google Scholar 

  • Weinreich M, Liang C, Stillman B (1999) The Cdc6p nucleotide-binding motif is required for loading Mcm proteins onto chromatin. Proc Natl Acad Sci U S A 96:441–446

    PubMed Central  CAS  PubMed  Google Scholar 

  • Wohlschlegel JA, Dhar SK, Prokhorova TA, Dutta A, Walter JC (2002) Xenopus Mcm10 binds to origins of DNA replication after Mcm2-7 and stimulates origin binding of Cdc45. Mol Cell 9:233–240

    CAS  PubMed  Google Scholar 

  • Xu X, Rochette PJ, Feyissa EA, Su TV, Liu Y (2009) MCM10 mediates RECQ4 association with MCM2-7 helicase complex during DNA replication. Embo J 28:3005–3014

    PubMed Central  CAS  PubMed  Google Scholar 

  • Yabuuchi H, Yamada Y, Uchida T, Sunathvanichkul T, Nakagawa T, Masukata H (2006) Ordered assembly of Sld3, GINS and Cdc45 is distinctly regulated by DDK and CDK for activation of replication origins. Embo J 25:4663–4674

    PubMed Central  CAS  PubMed  Google Scholar 

  • Yamada Y, Nakagawa T, Masukata H (2004) A novel intermediate in initiation complex assembly for fission yeast DNA replication. Mol Biol Cell 15:3740–3750. doi:10.1091/mbc.E04-04-0292

    PubMed Central  CAS  PubMed  Google Scholar 

  • Yardimci H, Loveland AB, Habuchi S, van Oijen AM, Walter JC (2010) Uncoupling of sister replisomes during eukaryotic DNA replication. Mol Cell 40:834–840. doi:10.1016/j.molcel.2010.11.027

    PubMed Central  CAS  PubMed  Google Scholar 

  • Yardimci H et al (2012) Bypass of a protein barrier by a replicative DNA helicase. Nature 492:205–209. doi:10.1038/nature11730

    PubMed Central  CAS  PubMed  Google Scholar 

  • Zachariae W, Schwab M, Nasmyth K, Seufert W (1998) Control of cyclin ubiquitination by CDK-regulated binding of Hct1 to the anaphase promoting complex. Science 282:1721–1724

    CAS  PubMed  Google Scholar 

  • Zegerman P, Diffley JF (2007) Phosphorylation of Sld2 and Sld3 by cyclin-dependent kinases promotes DNA replication in budding yeast. Nature 445:281–285

    CAS  PubMed  Google Scholar 

  • Zegerman P, Diffley JF (2010) Checkpoint-dependent inhibition of DNA replication initiation by Sld3 and Dbf4 phosphorylation. Nature 467:474–478

    PubMed Central  CAS  PubMed  Google Scholar 

  • Zhou Y, Wang TS (2004) A coordinated temporal interplay of nucleosome reorganization factor, sister chromatin cohesion factor, and DNA polymerase alpha facilitates DNA replication. Mol Cell Biol 24:9568–9579. doi:10.1128/MCB.24.21.9568-9579.2004

  • Zou L, Stillman B (1998) Formation of a preinitiation complex by S-phase cyclin CDK-dependent loading of Cdc45p onto chromatin. Science 280:593–596

    CAS  PubMed  Google Scholar 

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Acknowledgments

We would like to thank A. Okorokov, C. Herrera, Y. Javadi and F. Pisani for comments on the manuscript, and Imperial College and the MRC for funding.

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Tognetti, S., Riera, A. & Speck, C. Switch on the engine: how the eukaryotic replicative helicase MCM2–7 becomes activated. Chromosoma 124, 13–26 (2015). https://doi.org/10.1007/s00412-014-0489-2

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