Ctf4p facilitates Mcm10p to promote DNA replication in budding yeast

Wang, Jiafeng; Wu, Rentian; Lü, Yongjun; Liang, Chun

doi:10.1016/j.bbrc.2010.04.006

Cited by 26 publications

(32 citation statements)

References 35 publications

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“…Consistent with published data (Masumoto et al 2005;Wang et al 2010), we observed that Ctf4 was bound to chromatin mainly during S phase and dissociated in late S or G2 phase ( Figure 7A). Interestingly, chromatin dissociation of Ctf4 occurred concomitantly with the increase in H3K56ac ( Figure 7A).…”

Section: Ctf4 Association To Chromatin Is Not Notably Affected In Rrmsupporting

confidence: 92%

Replisome Function During Replicative Stress Is Modulated by Histone H3 Lysine 56 Acetylation Through Ctf4

et al. 2015

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Histone H3 lysine 56 acetylation in Saccharomyces cerevisiae is required for the maintenance of genome stability under normal conditions and upon DNA replication stress. Here we show that in the absence of H3 lysine 56 acetylation replisome components become deleterious when replication forks collapse at natural replication block sites. This lethality is not a direct consequence of chromatin assembly defects during replication fork progression. Rather, our genetic analyses suggest that in the presence of replicative stress H3 lysine 56 acetylation uncouples the Cdc45-Mcm2-7-GINS DNA helicase complex and DNA polymerases through the replisome component Ctf4. In addition, we discovered that the N-terminal domain of Ctf4, necessary for the interaction of Ctf4 with Mms22, an adaptor protein of the Rtt101-Mms1 E3 ubiquitin ligase, is required for the function of the H3 lysine 56 acetylation pathway, suggesting that replicative stress promotes the interaction between Ctf4 and Mms22. Taken together, our results indicate that Ctf4 is an essential member of the H3 lysine 56 acetylation pathway and provide novel mechanistic insights into understanding the role of H3 lysine 56 acetylation in maintaining genome stability upon replication stress. KEYWORDS Ctf4; H3K56 acetylation; Mms22; replicative stress; replisome T HE eukaryotic replisome consists of polymerases and an essential DNA helicase that are linked by a number of factors assembled during the initiation of chromosome replication. Progression of the replication fork depends on the activity of the replisome progression complex (RPC). This complex is uniquely present during S phase (Gambus et al. 2006) and remains associated with the replication fork until completion of DNA replication. In Saccharomyces cerevisiae, the RPC is made up of Mcm10, Mrc1, Tof1, Csm3, Ctf4, Top1, FACT (Spt16 and Pob3), and the CMG complex comprising Cdc45, , and the go ichi ni san (GINS) complex. The CMG constitutes the core replicative helicase responsible for the movement and activities of the replication fork (Pacek et al. 2006;Bochman and Schwacha 2009).The link between helicase and polymerases is a crucial determinant for the regulation of the replisome. The leadingstrand DNA polymerase-ɛ was recently shown to be integrated into the replisome via an interaction with the GINS complex (Sengupta et al. 2013). Furthermore, the DNA polymerasea-primase complex, which initiates DNA synthesis at replication origins and continues to prime Okazaki fragments at the fork, remains associated with the RPC via the Ctf4 trimer, which simultaneously interacts with the GINS complex (Gambus et al. 2009;Tanaka et al. 2009;Gosnell and Christensen 2011;Simon et al. 2014).Cells have evolved different mechanisms to maintain genome integrity under the conditions threatening replication progression (Jossen and Bermejo 2013;Leman and Noguchi 2013). The S-phase checkpoint mediated by MRC1 was initially characterized as a pathway activated by fork stalling and able to both stabilize the replisome and dela...

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Section: Ctf4 Association To Chromatin Is Not Notably Affected In Rrmsupporting

confidence: 92%

Replisome Function During Replicative Stress Is Modulated by Histone H3 Lysine 56 Acetylation Through Ctf4

et al. 2015

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“…p1ARS contains one replication origin ARS1, and p8ARSs carries seven additional copies of H4-ARS inserted into p1ARS. It has been well documented that all known replication initiation mutants have a higher rate of p1ARS loss compared with p8ARSs (Hogan and Koshland, 1992;Loo et al, 1995;Zhang et al, 2002;Cheng et al, 2010;Ma et al, 2010;Wang et al, 2010;Zhai et al, 2010). In control cells containing the empty BD vector, the loss rates of p1ARS and p8ARSs were 1.2% and 0.5% per generation, respectively (Fig.…”

Section: Cdt1p and Mcm6p Interact Through Their C-terminal Domainsmentioning

confidence: 81%

“…Matchmaker system III from Clontech was used for the yeast two-hybrid assay, conducted as previously described (Kan et al, 2008;Lai et al, 2011;Wang et al, 2010).…”

Section: Yeast Two-hybrid Assaymentioning

confidence: 99%

Cdt1p, through its interaction with Mcm6p, is required for the formation, nuclear accumulation and chromatin loading of the MCM complex

Wang

Liang

2012

Journal of Cell Science

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SummaryRegulation of DNA replication initiation is essential for the faithful inheritance of genetic information. Replication initiation is a multi-step process involving many factors including ORC, Cdt1p, Mcm2-7p and other proteins that bind to replication origins to form a pre-replicative complex (pre-RC). As a prerequisite for pre-RC assembly, Cdt1p and the Mcm2-7p heterohexameric complex accumulate in the nucleus in G1 phase in an interdependent manner in budding yeast. However, the nature of this interdependence is not clear, nor is it known whether Cdt1p is required for the assembly of the MCM complex. In this study, we provide the first evidence that Cdt1p, through its interaction with Mcm6p with the C-terminal regions of the two proteins, is crucial for the formation of the MCM complex in both the cytoplasm and nucleoplasm. We demonstrate that disruption of the interaction between Cdt1p and Mcm6p prevents the formation of the MCM complex, excludes Mcm2-7p from the nucleus, and inhibits pre-RC assembly and DNA replication. Our findings suggest a function for Cdt1p in promoting the assembly of the MCM complex and maintaining its integrity by interacting with Mcm6p.

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“…Therefore, we employed these plasmids to identify mutants defective in DNA replication initiation. Among many mutants in known and unknown replication initiation proteins (37,38), 4 an adk1…”

Section: Adk1mentioning

confidence: 99%

“…From a random ethane methyl sulfonate mutagenesis followed by a phenotypic screen (which we called the initiation of DNA replication, or IDR, screen) to identify genes involved in or regulating replication initiation in S. cerevisiae (37,38), 4 we isolated an adk1 G20S mutant that loses a single-ARS plasmid at a high rate and a multiple-ARS plasmid at a reduced rate. We show that both the adk1-td and adk1 G20S mutants have replication initiation defects, suggesting that Adk1p plays an important role in DNA replication initiation.…”

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confidence: 99%

ATP-dependent Pre-replicative Complex Assembly Is Facilitated by Adk1p in Budding Yeast

Cheng

Wang

et al. 2010

Journal of Biological Chemistry

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Pre-replicative complex (pre-RC) assembly is a critical part of the mechanism that controls the initiation of DNA replication, and ATP binding and hydrolysis by multiple pre-RC proteins are essential for pre-RC assembly and activation. Here, we demonstrate that Adk1p (adenylate kinase 1 protein) plays an important role in pre-RC assembly in Saccharomyces cerevisiae. Isolated from a genetic screen, adk1 G20S cells with a mutation within the nucleotide-binding site were defective in replication initiation. adk1⌬ cells were viable at 25°C but not at 37°C. Flow cytometry indicated that both the adk1-td (temperature-inducible degron) and adk1 G20S mutants were defective in S phase entry. Furthermore, Adk1p bound to chromatin throughout the cell cycle and physically interacted with Orc3p, whereas the Adk1 G20S protein had a reduced ability to bind chromatin and Orc3p without affecting the cellular ATP level. In addition, Adk1p associated with replication origins by ChIP assay. Finally, Adk1-td protein depletion prevented pre-RC assembly during the M-to-G 1 transition. We suggest that Adk1p regulates ATP metabolism on pre-RC proteins to promote pre-RC assembly and activation.The initiation of DNA replication governs the genome duplication in eukaryotes, which occurs once and only once per cell cycle. In Saccharomyces cerevisiae, replication initiation is critically controlled by pre-replicative complex (pre-RC) 3 assembly, which includes sequential loading of origin recognition complex (ORC) (1-3), Noc3p (4), Cdc6p (5-8), Cdt1p (9), and minichromosome maintenance (MCM) proteins (Mcm2-7p) (10, 11) onto autonomously replicating sequence (ARS) elements. In addition, ATP binding to and hydrolysis by Cdc6p and several subunits of ORC are essential for pre-RC assembly (12-15). At least 10 components (Orc1p, Orc4p, Orc5p, Cdc6p, and Mcm2-7p) of pre-RCs are ATPases (16) containing the Walker A and B motifs required for ATP binding and hydrolysis.Mutation analysis showed that all conserved ATP-binding motifs of these proteins are essential for cell viability (12,17,18).There have been numerous reports on the roles of binding and hydrolysis of ATP in pre-RC assembly. In S. cerevisiae, ATP binding of Orc1p activates ORC assembly and ORC binding to replication origins (1). Mutation within the Orc1p ATP-binding site prevents ORC binding to DNA in vitro and is lethal in vivo (19). Other studies indicated that mutations in the Walker A motif eliminate the ATP-binding and hydrolysis activities of Orc1p (20,21). When ATP binds to Orc1p, an initial round of chromatin loading of MCM proteins is permitted, whereas ATP hydrolysis is required for other rounds of MCM loading in vitro (13,22). Human ORC assembly in vitro is dependent on ATP binding and impaired by mutations in Orc4p or Orc5p ATP-binding sites (23,24). In S. cerevisiae, Cdc6p binding to ORC, which is dependent on the Cdc6p ATPase activity, changes the ORC structure and contributes to pre-RC assembly (25). Cdc6p mutated in the Walker B motif cannot interact with Orc1p in vitro ...

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Ctf4p facilitates Mcm10p to promote DNA replication in budding yeast

Cited by 26 publications

References 35 publications

Replisome Function During Replicative Stress Is Modulated by Histone H3 Lysine 56 Acetylation Through Ctf4

Replisome Function During Replicative Stress Is Modulated by Histone H3 Lysine 56 Acetylation Through Ctf4

Cdt1p, through its interaction with Mcm6p, is required for the formation, nuclear accumulation and chromatin loading of the MCM complex

ATP-dependent Pre-replicative Complex Assembly Is Facilitated by Adk1p in Budding Yeast

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