ATR Homolog Mec1 Controls Association of DNA Polymerase ζ-Rev1 Complex with Regions near a Double-Strand Break

Hirano, Yukinori; Sugimoto, Katsunori

doi:10.1016/j.cub.2006.01.063

Cited by 74 publications

(94 citation statements)

References 32 publications

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“…Immunoblotting analysis and viability assay after HO-induced DNA breaks were performed as described previously (Wakayama et al, 2001;Hirano and Sugimoto, 2006).…”

Section: Other Methodsmentioning

confidence: 99%

“…Chromatin immunoprecipitation was performed using anti-hemagglutinin (HA) (16B2) or anti-myc (9E10) antibodies as described previously (Hirano and Sugimoto, 2006). The PCR reaction was performed under nonsaturating conditions, in which the rate of PCR amplification was proportional to substrate concentration and cycling.…”

Section: Chromatin Immunoprecipitation (Chip) Assaymentioning

confidence: 99%

“…The cdc13-1 and stn1-13 strains were obtained from M. Charbonneau (Ecole Normale Superìeure de Lyon, Lyon, France) (Grandin et al, 2001). Epitope tagging of EXO1 and MRE11 was performed by a PCR-based strategy (Knop et al, 1999;Hirano and Sugimoto, 2006). The CDC13-myc strain was constructed using the integration plasmid as described previously (Taggart et al, 2002).…”

Section: Plasmids and Strainsmentioning

confidence: 99%

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Cdc13 Telomere Capping Decreases Mec1 Association but Does Not Affect Tel1 Association with DNA Ends

Hirano

Sugimoto

2007

MBoC

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Chromosome ends, known as telomeres, have to be distinguished from DNA breaks that activate DNA damage checkpoint. Two large protein kinases, ataxia-teleangiectasia mutated (ATM) and ATM-Rad3-related (ATR), control not only checkpoint activation but also telomere length. In budding yeast, Mec1 and Tel1 correspond to ATR and ATM, respectively. Here, we show that Cdc13-dependent telomere capping attenuates Mec1 association with DNA ends. The telomeric TG repeat sequence inhibits DNA degradation and decreases Mec1 accumulation at the DNA end. The TG-mediated degradation block requires binding of multiple Cdc13 proteins. The Mre11-Rad50-Xrs2 complex and Exo1 contribute to DNA degradation at DNA ends. Although the TG sequence impedes Exo1 association with DNA ends, it allows Mre11 association. Moreover, the TG sequence does not affect Tel1 association with the DNA end. Our results suggest that the Cdc13 telomere cap coordinates Mec1 and Tel1 accumulation rather than simply covering the DNA ends at telomeres. INTRODUCTIONDNA double-strand breaks (DSBs) are induced by exogenous DNA-damaging agents and carcinogens or by endogenous by-products, including reactive oxygen species (Friedberg et al., 2006). The repair of DSBs is crucial for maintaining genome stability. All organisms respond to DSBs by promptly launching the DNA damage response. This response involves the recruitment of DNA repair factors and the activation of signal transduction pathways, often termed DNA damage checkpoint pathways (Zhou and Elledge, 2000). Activation of the checkpoint pathway induces cell cycle arrest and expression of genes required for DNA repair.The checkpoint signals are initiated through two large protein kinases, ataxia-teleangiectasia mutated (ATM) and ATM-Rad3-related (ATR) (Zhou and Elledge, 2000;Abraham, 2001). ATM and ATR are highly conserved among eukaryotes. ATR is closely related to Mec1 in the budding yeast Saccharomyces cerevisiae and Rad3 in the fission yeast Schizosaccharomyces pombe. ATM homologues are termed Tel1 in both budding and fission yeasts. Current evidence indicates that the Mre11-Rad50 -Nbs1 (Xrs2 in budding yeast) complex is the primary sensor that recruits ATR/ Mec1 and ATM/Tel1 to DSBs (Nakada et al., 2003a(Nakada et al., , 2004Falck et al., 2005;You et al., 2005). In budding yeast, there is compelling evidence that the Mre11-Rad50 -Xrs2 (MRX) complex collaborates with exonuclease 1 (Exo1) in the generation of single-stranded DNA (ssDNA) at DSB ends (Krogh and Symington, 2004). ATM/Tel1 interacts with the C terminus of Nbs1/Xrs2 to localize to DNA ends (Nakada et al., 2003a;Falck et al., 2005;You et al., 2005). Meanwhile, the ssDNA that is generated at the DSB is covered with replication protein A (RPA) (Wold, 1997;Krogh and Symington, 2004). RPA-covered DNA recruits ATR/Mec1 to a region near the DSB end (Zou and Elledge, 2003;Nakada et al., 2005). ATM and ATR activate the downstream kinases CHK1 and CHK2 with assistance of checkpoint mediators, including 53BP1, BRCA1, and MDC1 (Zhou and Elledge, 2000;Bakke...

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“…Immunoblotting analysis and viability assay after HO-induced DNA breaks were performed as described previously (Wakayama et al, 2001;Hirano and Sugimoto, 2006).…”

Section: Other Methodsmentioning

confidence: 99%

Section: Chromatin Immunoprecipitation (Chip) Assaymentioning

confidence: 99%

Section: Plasmids and Strainsmentioning

confidence: 99%

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Cdc13 Telomere Capping Decreases Mec1 Association but Does Not Affect Tel1 Association with DNA Ends

Hirano

Sugimoto

2007

MBoC

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“…For instance, Rad9 of the S. pombe 9-1-1 complex, which forms a PCNA-like heterotrimeric clamp, associates with Mms2, and a mutant form of Rad9 incapable of interaction promotes mutagenesis in a TLS-dependent manner [133]. In budding yeast the phosphorylation by protein kinase Mec1 induces the re-localization of Rev1 and Polξ to sites of DNA double-strand breaks independently of mono-Ub PCNA [134]. Furthermore, the budding yeast 9-1-1 clamp physically interacts with the Rev7 subunit of Polζ and is partially required for spontaneous mutagenesis in a Polζ-dependent manner [135].…”

Section: Regulated Access Of Y-family Polymerases To the Damage Sitementioning

confidence: 99%

Eukaryotic DNA damage tolerance and translesion synthesis through covalent modifications of PCNA

Andersen¹,

Xiao³

2007

Cell Res

183

186

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npg In addition to well-defined DNA repair pathways, all living organisms have evolved mechanisms to avoid cell death caused by replication fork collapse at a site where replication is blocked due to disruptive covalent modifications of DNA. The term DNA damage tolerance (DDT) has been employed loosely to include a collection of mechanisms by which cells survive replication-blocking lesions with or without associated genomic instability. Recent genetic analyses indicate that DDT in eukaryotes, from yeast to human, consists of two parallel pathways with one being error-free and another highly mutagenic. Interestingly, in budding yeast, these two pathways are mediated by sequential modifications of the proliferating cell nuclear antigen (PCNA) by two ubiquitination complexes Rad6-Rad18 and Mms2-Ubc13-Rad5. Damage-induced monoubiquitination of PCNA by Rad6-Rad18 promotes translesion synthesis (TLS) with increased mutagenesis, while subsequent polyubiquitination of PCNA at the same K164 residue by Mms2-Ubc13-Rad5 promotes error-free lesion bypass. Data obtained from recent studies suggest that the above mechanisms are conserved in higher eukaryotes. In particular, mammals contain multiple specialized TLS polymerases. Defects in one of the TLS polymerases have been linked to genomic instability and cancer. DNA damage toleranceIn the presence of spontaneous or carcinogen-induced DNA damage, living cells have to maintain and complete DNA synthesis or risk replication fork collapse. Since the process of DNA licensing is to ensure the genome is duplicated once and only once during each cell cycle, stalled or collapsed replication forks may not be able to restart, which often results in double-strand breaks (DSBs) and causes compromised genome integrity or cell death. In addition to highly conserved DNA repair pathways, all living organisms have evolved schemes to ensure continuation of DNA synthesis in the presence of damage. These schemes were originally termed DNA postreplication repair (PRR) due to observations of transient shortened nascent DNA structures following S phase in response to DNA damage. In bacteria and unicellular yeast, these shortened DNA segments can be measured by an alkaline sedimentation assay [1] or directly observed in electron micrographs [2]. In wild-type cells, these truncated DNA segments were restored to full length following a short recovery time. One typical experiment [1] involved the restoration of the nascent strand following UV exposure in nucleotide excision repair (NER)-deficient cells and was originally assumed to represent a mechanism of DNA repair. However, further investigation revealed that, although the nascent fragments were re-annealed, the original UV-induced pyrimidine dimers, which were responsible for the generation of single-strand gaps, often persisted in the genome [3,4]. It was argued that the replication-blocking lesion was not necessarily corrected, but rather transiently bypassed and carried over to the next generation. Perhaps it is more beneficial for the organi...

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“…The Rev1 protein has both a dCMP transferase activity that may function during DNA damage bypass [18,19] and an important role as a scaffolding protein which associates with several translesion synthesis polymerases. Yeast pol ζ interacts with Rev1 as shown by co-immunoprecipitation [20,21] and this association enhances the efficiency of extension from mismatched primer-templates and AP sites [19,21]. Yeast DNA pol ζ also interacts functionally with PCNA and with the alternative "9-1-1" complex clamp [22, The leading (top) and lagging (bottom) strand of a DNA replication fork during S-phase is represented.…”

Section: Dna Pol ζ In Saccharomyces Cerevisiaementioning

confidence: 99%

DNA polymerase zeta (pol ζ) in higher eukaryotes

et al. 2007

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Most current knowledge about DNA polymerase zeta (pol ζ) comes from studies of the enzyme in the budding yeast Saccharomyces cerevisiae, where pol ζ consists of a complex of the catalytic subunit Rev3 with Rev7, which associates with Rev1. Most spontaneous and induced mutagenesis in yeast is dependent on these gene products, and yeast pol ζ can mediate translesion DNA synthesis past some adducts in DNA templates. Study of the homologous gene products in higher eukaryotes is in a relatively early stage, but additional functions for the eukaryotic proteins are already apparent. Suppression of vertebrate REV3L function not only reduces induced point mutagenesis but also causes larger-scale genome instability by raising the frequency of spontaneous chromosome translocations. Disruption of Rev3L function is tolerated in Drosophila, Arabidopsis, and in vertebrate cell lines under some conditions, but is incompatible with mouse embryonic development. Functions for REV3L and REV7(MAD2B) in higher eukaryotes have been suggested not only in translesion DNA synthesis but also in some forms of homologous recombination, repair of interstrand DNA crosslinks, somatic hypermutation of immunoglobulin genes and cell-cycle control. This review discusses recent developments in these areas.

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ATR Homolog Mec1 Controls Association of DNA Polymerase ζ-Rev1 Complex with Regions near a Double-Strand Break

Cited by 74 publications

References 32 publications

Cdc13 Telomere Capping Decreases Mec1 Association but Does Not Affect Tel1 Association with DNA Ends

Cdc13 Telomere Capping Decreases Mec1 Association but Does Not Affect Tel1 Association with DNA Ends

Eukaryotic DNA damage tolerance and translesion synthesis through covalent modifications of PCNA

DNA polymerase zeta (pol ζ) in higher eukaryotes

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