Yeast Rad17/Mec3/Ddc1: A sliding clamp for the DNA damage checkpoint

Majka, Jerzy; Burgers, Peter M.

doi:10.1073/pnas.0437148100

Cited by 244 publications

(243 citation statements)

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“…Rad17/3/1 and RFCRad24 interact in the absence of DNA (6,9,10). As previously observed with PCNA and RFC, the formation of a stable Rad17/ 3/1⅐RFC-Rad24 complex was strongly enhanced by ATP binding but did not require its hydrolysis (12).…”

supporting

confidence: 57%

“…As previously observed with PCNA and RFC, the formation of a stable Rad17/ 3/1⅐RFC-Rad24 complex was strongly enhanced by ATP binding but did not require its hydrolysis (12). Loading of the Rad17/3/1 clamp around partial duplex DNA can be mediated by either ATP or ATP␥S; however, ATP hydrolysis was required to release the clamp from the clamp loader and permit sliding of Rad17/3/1 across double-stranded DNA (8,10). The nature of the DNA substrate for this system has been a matter of discrepancy.…”

mentioning

confidence: 64%

“…The nature of the DNA substrate for this system has been a matter of discrepancy. We determined that yeast Rad17/3/1 is loaded onto partial duplex DNA with either 5Ј-or 3Ј-recessed termini and can even be loaded onto double-stranded DNA ends (10). However, loading on a normal template-primer substrate, i.e.…”

mentioning

confidence: 99%

“…The checkpoint clamp proteins Rad17, Mec3, and Ddc1 (Rad1, Hus1, and Rad9 in Schizosaccharomyces pombe and human) display sequence homology with each other and with PCNA, and this complex has been purified as a heterotrimer from several sources (4 -10). Similarly, the DNA damage checkpoint clamp loader consisting of Saccharomyces cerevisiae Rad24 (Rad17 in S. pombe and human) together with the four small Rfc2-5 subunits has been purified as a heteropentameric complex from several organisms (5)(6)(7)(8)(9)(10)(11). In this study, the checkpoint clamp is designated as Rad17/3/1 and the clamp loader is designated as RFC-Rad24.…”

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confidence: 99%

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Requirement for ATP by the DNA Damage Checkpoint Clamp Loader

Majka

Chung

Burgers

2004

Journal of Biological Chemistry

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The DNA damage clamp loader replication factor C (RFC-Rad24) consists of the Rad24 protein and the four small Rfc2-5 subunits of RFC. This complex loads the heterotrimeric DNA damage clamp consisting of Rad17, Mec3, and Ddc1 (Rad17/3/1) onto partial duplex DNA in an ATP-dependent manner. Interactions between the clamp loader and the clamp have been proposed to mirror those of the replication clamp loader RFC and the sliding clamp proliferating cell nuclear antigen (PCNA). In that system, three ATP molecules bound to the Rfc2, Rfc3, and Rfc4 subunits are necessary and sufficient for efficient loading of PCNA, whereas ATP binding to Rfc1 is not required. In contrast, in this study, we show that mutant RFC-Rad24 with a rad24-K115E mutation in the ATP-binding domain of Rad24 shows defects in the ATPase of the complex and is defective for interaction with Rad17/3/1 and for loading of the checkpoint clamp. A similar defect was measured with a mutant RFCRad24 clamp loader carrying a rfc4K55R ATP-binding mutation, whereas the rfc4K55E clamp loader showed partial loading activity, in agreement with genetic studies of these mutants. These studies show that ATP utilization by the checkpoint clamp/clamp loader system is effectively different from that by the structurally analogous replication system.

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confidence: 57%

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confidence: 64%

mentioning

confidence: 99%

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confidence: 99%

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Requirement for ATP by the DNA Damage Checkpoint Clamp Loader

Majka

Chung

Burgers

2004

Journal of Biological Chemistry

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“…Completion of the 5Ј-to-3Ј degradation could relocate checkpoint proteins, thereby attenuating checkpoint activation. The PCNA-like checkpoint clamps have been shown to target partial duplex DNA (Majka and Burgers, 2003;Majka et al, 2006). Although retained at early time points, the other strand of the distal DNA fragment disap- peared at 6 h after HO induction.…”

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confidence: 99%

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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DNADamage: Sensing

Yang

Zou

2008

Wiley Encyclopedia of Chemical Biology

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Maintenance of genomic stability is essential for the survival of cells, organisms, and species. Genomic stability relies on the complete and accurate transmission of genetic materials from mother cells to daughter cells and from one generation to the next. This task is daunting, however, because the genome is constantly challenged by numerous intrinsic and extrinsic stresses that damage deoxyribonucleic acid (DNA). If left untreated, such damage can destabilize the genome by introducing gene mutations, duplications, and chromosomal rearrangements such as deletions and inversions, which fortunately does not normally happen as cells evolve a complex damage sensing and signaling mechanism named the DNA damage and replication checkpoint. Checkpoint activation effectively pauses the progression of the cell cycle, allowing more time for the removal of DNA lesion. Moreover, activated checkpoint also regulates and coordinates a number of cellular processes including DNA repair, DNA replication, and chromatin remodeling to alleviate the stress on the genome. At the core of the checkpoint‐signaling network, a family of phosphoinositide‐3‐kinase (PI‐3K)‐like kinases and their regulatory partners serve as both DNA damage sensors and initiators of checkpoint signaling. In this article, we will discuss in detail how DNA damage is sensed and processed by the DNA damage and replication checkpoint. We will also discuss how signals generated at sites of DNA damage are propagated and relayed through the checkpoint pathways to influence other cellular events.

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Yeast Rad17/Mec3/Ddc1: A sliding clamp for the DNA damage checkpoint

Cited by 244 publications

References 39 publications

Requirement for ATP by the DNA Damage Checkpoint Clamp Loader

Requirement for ATP by the DNA Damage Checkpoint Clamp Loader

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

DNADamage: Sensing

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