Properties of the human Cdc45/Mcm2-7/GINS helicase complex and its action with DNA polymerase ε in rolling circle DNA synthesis

Kang, Young‐Hoon; Galal, Wiebke Chemnitz; Farina, Andrea; Tappin, Inger; Hurwitz, Jerard

doi:10.1073/pnas.1203734109

Cited by 133 publications

(174 citation statements)

References 26 publications

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“…The CMG complex is a highly processive and active helicase with high ATPase rates, whereas Mcm2-7 is a very poor ATPase, and particular reaction conditions are required to observe even weak activity in vitro (15)(16)(17). These data suggest that Cdc45 and GINS are required for full activation of the replication fork helicase (15,16).…”

mentioning

confidence: 80%

“…The CMG Complex May Be a Poor Helicase in Vitro because of the High Level of ssDNA Present in Biochemical Assays-Recent work by the Hurwitz group (17) demonstrates that the CMG helicase is processive only in the presence of a polymerase, RPA, or E. coli single-stranded binding protein. The data from this work suggest that the CMG complex may require E. coli SSB for processive helicase activity because singlestranded DNA present in these biochemical assays is toxic to the CMG.…”

Section: Discussionmentioning

confidence: 99%

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Cdc45 Protein-Single-stranded DNA Interaction Is Important for Stalling the Helicase during Replication Stress

Bruck

Kaplan

2013

Journal of Biological Chemistry

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Background: Polymerase stalling is coupled with helicase stalling at a eukaryotic replication fork by an unknown mechanism. Results: When a Cdc45-ssDNA binding mutant is expressed in budding yeast cells exposed to hydroxyurea, there is uncoupling of the helicase from the polymerase. Conclusion: Cdc45-ssDNA interaction is important during replication stress. Significance: A new model explains how polymerase stalling is coupled with helicase stalling.

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mentioning

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Cdc45 Protein-Single-stranded DNA Interaction Is Important for Stalling the Helicase during Replication Stress

Bruck

Kaplan

2013

Journal of Biological Chemistry

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“…This is supported by the adaptation of yPold3 for Okazaki fragment processing and also by recent advances in the study of the GINS complex, which is a component of the eukaryotic initiation helicase, the CMG complex [MacNeill, 2010;Bermudez et al, 2011;Costa et al, 2011;Kang et al, 2012]. The recruitment of Pol a and Pol e to the GINS complex is a key step in initiation of DNA replication, and the association of Pol e with the GINS complex provides for a separate structural unit associated with ongoing leading strand synthesis [Seki et al, 2006;Chang et al, 2007].…”

Section: Role Of Pol D At the Replication Forkmentioning

confidence: 93%

Regulation of human DNA polymerase delta in the cellular responses to DNA damage

Lee

Zhang

Lin

et al. 2012

Environ and Mol Mutagen

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The p12 subunit of polymerase delta (Pol d) is degraded in response to DNA damage induced by UV, alkylating agents, oxidative, and replication stresses. This leads to the conversion of the Pol d4 holoenzyme to the heterotrimer, Pol d3. We review studies that establish that Pol d3 formation is an event that could have a major impact on cellular processes in genomic surveillance, DNA replication, and DNA repair. p12 degradation is dependent on the apical ataxia telangiectasia and Rad3 related (ATR) kinase and is mediated by the ubiquitin-proteasome system. Pol d3 exhibits properties of an ''antimutator'' polymerase, suggesting that it could contribute to an increased surveillance against mutagenesis, for example, when Pol d carries out bypass synthesis past small base lesions that engage in spurious base pairing. Chromatin immunoprecipitation analysis and examination of the spatiotemporal recruitment of Pol d to sites of DNA damage show that Pol d3 is the primary form of Pol d associated with cyclobutane pyrimidine dimer lesions and therefore should be considered as the operative form of Pol d engaged in DNA repair. We propose a model for the switching of Pol d with translesion polymerases, incorporating the salient features of the recently determined structure of monoubiquitinated proliferating cell nuclear antigen and emphasizing the role of Pol d3. Because of the critical role of Pol d activity in DNA replication and repair, the formation of Pol d3 in response to DNA damage opens the prospect that pleiotropic effects may ensue. This opens the horizons for future exploration of how this novel response to DNA damage contributes to genomic stability. Environ. Mol. Mutagen. 53:683-698, 2012. V V C 2012 Wiley Periodicals, Inc.

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“…6, lanes 3-6). Previous studies in HeLa cells showed that the hCMG complex was found stably associated only in the chromatin fraction (14). Histone H3, which is localized mostly on chromatin, was used as a quality control of the lysate fractionation (Fig.…”

Section: Significancementioning

confidence: 99%

“…The CMG complex, composed of stoichiometric levels of Cdc45, Mcm2-7, and GINS, was isolated from Drosophila embryos, as well as insect cells, using the baculovirus-expression system and found to contain 3′→5′ DNA helicase activity (12,13). Subsequently, the hCMG complex was purified and shown to support the coupled helicase-Pol e-dependent synthesis of leading strand DNA in vitro (14).…”

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confidence: 99%

Interaction between human Ctf4 and the Cdc45/Mcm2-7/GINS (CMG) replicative helicase

Kang

Farina

Bermudez

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Chromosome transmission fidelity 4 (Ctf4) is a conserved protein required for DNA replication. In this report, interactions between human Ctf4 (hCtf4) and the replicative helicase containing the cell division cycle 45 (Cdc45)/minichromosome maintenance 2-7 (Mcm2-7)/Go, Ichi, Nii, and San (GINS) (CMG) proteins [human CMG (hCMG) complex] were examined. The hCtf4-CMG complex was isolated following in vitro interaction of purified proteins (hCtf4 plus the hCMG complex), coinfection of Spodoptera frugiperda (Sf9) insect cells with viruses expressing the hCMG complex and hCtf4, and from HeLa cell chromatin after benzonase and immunoprecipitation steps. The stability of the hCtf4-CMG complex depends upon interactions between hCtf4 and multiple components of the hCMG complex. The hCtf4-CMG complex, like the hCMG complex, contains DNA helicase activity that is more salt-resistant than the helicase activity of the hCMG complex. We demonstrate that the hCtf4-CMG complex contains a homodimeric hCtf4 and a monomeric hCMG complex and suggest that the homodimeric hCtf4 acts as a platform linking polymerase α to the hCMG complex. The role of the hCMG complex as the core of the replisome is also discussed.replisome core structure | dimerization | replication initiation E ukaryotic DNA replication is a stepwise process by which protein complexes are assembled on chromatin. During the G1 phase of the cell cycle, the origin recognition complex (ORC) binds to replication origins and recruits cell division cycle 6 (Cdc6) (1). This complex leads to the association of cdc10 dependent transcript 1 (Cdt1)/Mcm2-7 with chromatin and the loading of the Mcm2-7 complex as a head-to-head dimer (2, 3). At the G1/S transition, this prereplication complex is altered further by a number of replication initiation factors whose actions are facilitated by the cyclin-dependent and cell division cycle 7 (Cdc7)-dumbbell former 4 (Dbf4) kinases (4). These replication initiation factors [which include synthetically lethal with dpb11-1 (Sld)2, Sld3, Sld7, DNA polymerase B possible subunit 11 (Dpb11), and DNA polymerase e (Pol e) in budding yeast] play critical roles resulting in the interaction of Cdc45 and GINS with the Mcm2-7 complex and the formation of the replicative DNA helicase complex containing Cdc45/Mcm2-7/GINS (CMG) (5). Other replication factors (including Mcm10 and Ctf4) contribute to the activation of the CMG helicase and association of proteins that recruit the replicative Pols to effect DNA replication (6, 7). Interactions between TopBP1-interacting, replication-stimulating protein (Treslin)

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Properties of the human Cdc45/Mcm2-7/GINS helicase complex and its action with DNA polymerase ε in rolling circle DNA synthesis

Cited by 133 publications

References 26 publications

Cdc45 Protein-Single-stranded DNA Interaction Is Important for Stalling the Helicase during Replication Stress

Cdc45 Protein-Single-stranded DNA Interaction Is Important for Stalling the Helicase during Replication Stress

Regulation of human DNA polymerase delta in the cellular responses to DNA damage

Interaction between human Ctf4 and the Cdc45/Mcm2-7/GINS (CMG) replicative helicase

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