cdc7-1 A temperature sensitive cell-cycle mutant which interferes with induced mutagenesis in Saccharomyces cerevisiae

Njagi, G. D. E.; Kilbey, Brian J.

doi:10.1007/bf00337951

Cited by 60 publications

(31 citation statements)

References 12 publications

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“…CDC7 may be involved in the initiation of mitotic DNA synthesis, premeiotic recombination, formation ofascospores, and error-prone DNA repair (22,23 According to indirect immunofluorescence and subcellular fractionation studies, both forms of CDC7 are concentrated in the nuclei of CDC7-overproducing cells. We are not sure whether the staining patterns reflect a role for-CDC7 in both the cytoplasm and nucleus or if they simply result from the delocalization of the exclusively nuclear protein due to overproduction.…”

Section: Discussionmentioning

confidence: 99%

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

Yoon

Campbell

1991

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

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The CDC7 protein of Saccharomyces cerevisiae may be involved in the G1/S-phase transition and/or in the initiation of mitotic DNA synthesis. The CDC7 gene has two in-frame AUG codons as possible translation start sites, which would produce 58-and 56-kDa proteins, respectively. Both p58 and p56 derived from recombinant plasmids complement the temperature-sensitive growth defect of the cdc7-1 allele. To determine the biochemical function of the CDC7 protein, the CDC7 gene was cloned and polyclonal antibodies were produced against the CDC7 protein. CDC7 immune complexes prepared from yeast with these antibodies phosphorylate histone H1. Kinase activity is thermolabile in strains carrying the cdc7-1 temperature-sensitive mutant allele and is elevated >10-fold in strains carrying plasmids overexpressing either p56 or p58, confirming that the kinase in the immunoprecipitates is the CDC7 gene product. In addition, we show that CDC7 is a phosphoprotein itself. Indirect immunofluorescence and biochemical fractionation show that the CDC7 protein is present at relatively high concentrations in the nucleus compared with the cytoplasm, suggesting that nuclear proteins may be substrates for the CDC7 protein.Two major events define the eukaryotic cell cycle: replication of the chromosomes during S phase and segregation of the chromosomes during mitosis. Replication and segregation are separated by two gap periods, G1 and G2, during which the cells prepare for these events. There are also two major control points for the cell cycle in G1 and G2, the point of commitment to DNA synthesis in G1 and the regulation of progression into mitosis in G2. Although we have made great strides in understanding mitotic control in the past three years, relatively little is known about the events in G1 that lead to S phase. In yeast, the point of commitment to DNA synthesis is defined as "Start", early in G1. DNA synthesis does not ensue immediately after Start, however, and it is not clear whether the lag is due to assembly of the replication apparatus or to a cascade of controls initiated at Start. CDC28, CDC4, and CDC7 define a dependent series of events set in motion at Start (1-3). CDC28 encodes a protein kinase subunit, the analog of the highly conserved cdc2+/ MPF kinase subunit involved in regulation of mitosis (3).CDC4 is homologous to one subunit of the signal transducing protein, transducin (4). The DNA sequence of the CDC7 gene predicts a protein that has homology to catalytic domains of the protein kinases CDC2/CDC28, NIM1, and a number of kinase-related transforming proteins (5). The CDC7 sequence differs from that of other protein kinases in the so-called phosphate acceptor domain, however (5, 6). We have been interested in the role of CDC7 in the events linking Start and the initiation of DNA replication. Mutations in CDC7 appear to block a precondition for DNA synthesis because cells carrying these lesions cannot start new rounds of DNA replication after a shift from permissive to nonpermissive temperature (7,8). The cd...

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Section: Discussionmentioning

confidence: 99%

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

Yoon

Campbell

1991

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

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“…Consistent with this idea, Cdc7-Dbf4 has been implicated in error-prone TLS. Cdc7 is required for induced mutagenesis by UV and MMS, and combined mutations of CDC7 and RAD5 cause synergistic increases in UV and MMS sensitivities (Njagi and Kilbey 1982;Pessoa-Brandao and Sclafani 2004). This indicates that Rad5 and Cdc7-Dbf4-dependent TLS function in parallel and suggests that Rad53 may regulate this choice (Fig.…”

Section: Rad53 Regulates Replication Fork Restart Genes and Developmentmentioning

confidence: 92%

Rad53 regulates replication fork restart after DNA damage in Saccharomyces cerevisiae

Szyjka¹,

Aparicio²,

Viggiani³

et al. 2008

Genes Dev.

103

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Replication fork stalling at a DNA lesion generates a damage signal that activates the Rad53 kinase, which plays a vital role in survival by stabilizing stalled replication forks. However, evidence that Rad53 directly modulates the activity of replication forks has been lacking, and the nature of fork stabilization has remained unclear. Recently, cells lacking the Psy2-Pph3 phosphatase were shown to be defective in dephosphorylation of Rad53 as well as replication fork restart after DNA damage, suggesting a mechanistic link between Rad53 deactivation and fork restart. To test this possibility we examined the progression of replication forks in methyl-methanesulfonate (MMS)-damaged cells, under different conditions of Rad53 activity. Hyperactivity of Rad53 in pph3⌬ cells slows fork progression in MMS, whereas deactivation of Rad53, through expression of dominant-negative Rad53-KD, is sufficient to allow fork restart during recovery. Furthermore, combined deletion of PPH3 and PTC2, a second, unrelated Rad53 phosphatase, results in complete replication fork arrest and lethality in MMS, demonstrating that Rad53 deactivation is a key mechanism controlling fork restart. We propose a model for regulation of replication fork progression through damaged DNA involving a cycle of Rad53 activation and deactivation that coordinates replication restart with DNA repair.[Keywords: DNA replication fork; DNA damage; DNA repair; cell cycle checkpoint; BrdU; phosphatase; microarray] Supplemental material is available at http://www.genesdev.org.

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“…Indeed, at least in yeast, the Cdc7 kinase has already been implicated in DNA repair and mutagenesis. Thus, mutants of cdc7 exhibit varying degrees of mutability in response to genotoxic insults, including UV light, ethylmethanesulfonate, nitrosoguanidine, and nitrogen mustard (Njagi and Kilbey 1982). Interestingly, CDC7 belongs to the RAD6 epistasis group (Njagi and Kilbey 1982;PessoaBrandao and Sclafani 2004) of genes that control a relatively poorly understood DNA repair pathway of translesion DNA synthesis and damage tolerance.…”

mentioning

confidence: 99%

ATR–Chk1–APC/C^Cdh1-dependent stabilization of Cdc7–ASK (Dbf4) kinase is required for DNA lesion bypass under replication stress

et al. 2013

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Cdc7 kinase regulates DNA replication. However, its role in DNA repair and recombination is poorly understood. Here we describe a pathway that stabilizes the human Cdc7-ASK (activator of S-phase kinase; also called Dbf4), its regulation, and its function in cellular responses to compromised DNA replication. Stalled DNA replication evoked stabilization of the Cdc7-ASK (Dbf4) complex in a manner dependent on ATR-Chk1-mediated checkpoint signaling and its interplay with the anaphase-promoting complex/cyclosome Cdh1 (APC/C Cdh1 ) ubiquitin ligase. Mechanistically, Chk1 kinase inactivates APC/C Cdh1 through degradation of Cdh1 upon replication block, thereby stabilizing APC/C Cdh1 substrates, including Cdc7-ASK (Dbf4). Furthermore, motif C of ASK (Dbf4) interacts with the N-terminal region of RAD18 ubiquitin ligase, and this interaction is required for chromatin binding of RAD18. Impaired interaction of ASK (Dbf4) with RAD18 disables foci formation by RAD18 and hinders chromatin loading of translesion DNA polymerase h. These findings define a novel mechanism that orchestrates replication checkpoint signaling and ubiquitin-proteasome machinery with the DNA damage bypass pathway to guard against replication collapse under conditions of replication stress.

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cdc7-1 A temperature sensitive cell-cycle mutant which interferes with induced mutagenesis in Saccharomyces cerevisiae

Cited by 60 publications

References 12 publications

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

Rad53 regulates replication fork restart after DNA damage in Saccharomyces cerevisiae

ATR–Chk1–APC/C^Cdh1-dependent stabilization of Cdc7–ASK (Dbf4) kinase is required for DNA lesion bypass under replication stress

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cdc7-1 A temperature sensitive cell-cycle mutant which interferes with induced mutagenesis in Saccharomyces cerevisiae

Cited by 60 publications

References 12 publications

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

Rad53 regulates replication fork restart after DNA damage in Saccharomyces cerevisiae

ATR–Chk1–APC/CCdh1-dependent stabilization of Cdc7–ASK (Dbf4) kinase is required for DNA lesion bypass under replication stress

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ATR–Chk1–APC/C^Cdh1-dependent stabilization of Cdc7–ASK (Dbf4) kinase is required for DNA lesion bypass under replication stress