Histone methyltransferase Dot1 and Rad9 inhibit single-stranded DNA accumulation at DSBs and uncapped telomeres

Lazzaro, Federico; Sapountzi, Vasileia; Granata, Magda; Pellicioli, Achille; Vaze, Moreshwar B.; Haber, James E.; Plevani, Paolo; Lydall, David; Muzi-Falconi, Marco

doi:10.1038/emboj.2008.81

Cited by 147 publications

(202 citation statements)

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“…Therefore, the relationship between chromatin and the telomere cap is complex. Recent experiments show how the histone H3K79 methyl transferase Dot1 (identifed as UD S in our study) is required for checkpoint activation and inhibition of resection in cdc13-1 mutants (Lazzaro et al 2008).…”

Section: Discussionmentioning

confidence: 52%

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A Genomewide Suppressor and Enhancer Analysis of cdc13-1 Reveals Varied Cellular Processes Influencing Telomere Capping in Saccharomyces cerevisiae

Addinall

Downey

et al. 2008

Genetics

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In Saccharomyces cerevisiae, Cdc13 binds telomeric DNA to recruit telomerase and to ''cap'' chromosome ends. In temperature-sensitive cdc13-1 mutants telomeric DNA is degraded and cell-cycle progression is inhibited. To identify novel proteins and pathways that cap telomeres, or that respond to uncapped telomeres, we combined cdc13-1 with the yeast gene deletion collection and used high-throughput spot-test assays to measure growth. We identified 369 gene deletions, in eight different phenotypic classes, that reproducibly demonstrated subtle genetic interactions with the cdc13-1 mutation. As expected, we identified DNA damage checkpoint, nonsense-mediated decay and telomerase components in our screen. However, we also identified genes affecting casein kinase II activity, cell polarity, mRNA degradation, mitochondrial function, phosphate transport, iron transport, protein degradation, and other functions. We also identified a number of genes of previously unknown function that we term RTC, for restriction of telomere capping, or MTC, for maintenance of telomere capping. It seems likely that many of the newly identified pathways/ processes that affect growth of budding yeast cdc13-1 mutants will play evolutionarily conserved roles at telomeres. The high-throughput spot-testing approach that we describe is generally applicable and could aid in understanding other aspects of eukaryotic cell biology.

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Section: Discussionmentioning

confidence: 52%

“…help recruit Rad9 to chromatin and, like Rad9, inhibits resection at uncapped telomeres (Lazzaro et al 2008).…”

Section: Discussionmentioning

confidence: 99%

A Genomewide Suppressor and Enhancer Analysis of cdc13-1 Reveals Varied Cellular Processes Influencing Telomere Capping in Saccharomyces cerevisiae

Addinall

Downey

et al. 2008

Genetics

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“…A trivial explanation for this would be that there was incomplete inhibition of Cdc28-as1, despite using a high concentration of 1-NM-PP1. A more speculative interpretation would be that telomere chromatin is disrupted in the ten1-105 strain such that some Cdk1-independent resection occurs, as has been shown at a DSB in strains that are deficient for dot1, a methyltransferase, or for rad9, a checkpoint mediator that binds to modified chromatin (Lazzaro et al 2008).…”

Section: Discussionmentioning

confidence: 99%

TEN1Is Essential forCDC13-Mediated Telomere Capping

Petreaca

Gasparyan

et al. 2009

Genetics

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Telomere binding proteins protect chromosome ends from degradation and mask chromosome termini from checkpoint surveillance. In Saccharomyces cerevisiae, Cdc13 binds single-stranded G-rich telomere repeats, maintaining telomere integrity and length. Two additional proteins, Ten1 and Stn1, interact with Cdc13 but their contributions to telomere integrity are not well defined. Ten1 is known to prevent accumulation of aberrant single-stranded telomere DNA; whether this results from defective end protection or defective telomere replication is unclear. Here we report our analysis of a new group of ten1 temperature-sensitive (ts) mutants. At permissive temperatures, ten1-ts strains display greatly elongated telomeres. After shift to nonpermissive conditions, however, ten1-ts mutants accumulate extensive telomeric single-stranded DNA. Cdk1 activity is required to generate these single-stranded regions, and deleting the EXO1 nuclease partially suppresses ten1-ts growth defects. This is similar to cdc13-1 mutants, suggesting ten1-ts strains are defective for end protection. Moreover, like Cdc13, our analysis reveals Ten1 promotes de novo telomere addition. Interestingly, in ten1-ts strains at high temperatures, telomeric singlestranded DNA and Rad52-YFP repair foci are strongly induced despite Cdc13 remaining associated with telomeres, revealing Cdc13 telomere binding is not sufficient for end protection. Finally, unlike cdc13-1 mutants, ten1-ts strains display strong synthetic interactions with mutations in the POLa complex. These results emphasize that Cdc13 relies on Ten1 to execute its essential function, but leave open the possibility that Ten1 has a Cdc13-independent role in DNA replication.

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“…The DNA damage checkpoint has been shown to be required for efficient DSB repair where Rad24 Sc is required for resection and, thus, ssDNA production (Aylon and Kupiec 2003), while Rad9 Sc has been shown to limit ssDNA production associated with DSB repair (Lazzaro et al 2008). This function appears to be conserved, as the human Rad9 homolog 53BP1 has been shown to block extensive resection and suppress the repair defect of BRCA1 (Bunting et al 2010).…”

Section: Checkpoint-dependent Ssdna Regulation and Dsb Repairmentioning

confidence: 99%

Break-induced ATR and Ddb1–Cul4^Cdt2 ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

Moss

Tinline-Purvis²,

Walker³

et al. 2010

Genes Dev.

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Nucleotide synthesis is a universal response to DNA damage, but how this response facilitates DNA repair and cell survival is unclear. Here we establish a role for DNA damage-induced nucleotide synthesis in homologous recombination (HR) repair in fission yeast. Using a genetic screen, we found the Ddb1-Cul4 Cdt2 ubiquitin ligase complex and ribonucleotide reductase (RNR) to be required for HR repair of a DNA double-strand break (DSB). The Ddb1-Cul4 Cdt2 ubiquitin ligase complex is required for degradation of Spd1, an inhibitor of RNR in fission yeast. Accordingly, deleting spd1 + suppressed the DNA damage sensitivity and the reduced HR efficiency associated with loss of ddb1 + or cdt2 + . Furthermore, we demonstrate a role for nucleotide synthesis in postsynaptic gap filling of resected ssDNA ends during HR repair. Finally, we define a role for Rad3 (ATR) in nucleotide synthesis and HR through increasing Cdt2 nuclear levels in response to DNA damage. Our findings support a model in which breakinduced Rad3 and Ddb1-Cul4 Cdt2 ubiquitin ligase-dependent Spd1 degradation and RNR activation promotes postsynaptic ssDNA gap filling during HR repair.[Keywords: Rad3; Ddb1-Cul4 Cdt2 ubiquitin ligase; Spd1; ribonucleotide reductase; homologous recombination repair; fission yeast] Supplemental material is available at http://www.genesdev.org. Received July 16, 2010; revised version accepted October 15, 2010. DNA double-strand breaks (DSBs) are potentially lethal lesions that, if left undetected or repaired incorrectly, can threaten the integrity of the genome. DSBs arise at a low frequency during normal cell metabolism and can also arise from exposure to DNA-damaging agents such as ionizing radiation (IR) (Shrivastav et al. 2008), potentially leading to chromosomal rearrangements, cancer, or cell death (Pfeiffer et al. 2000). Consequently, an intricate network of cellular responses for detection and accurate repair of such lesions exists within the cell (Jackson and Bartek 2009).Cells have evolved two distinct repair pathways to maintain genome integrity following a DSB: nonhomologous end-joining (NHEJ), in which DNA ends are directly ligated, and homologous recombination (HR) (Shrivastav et al. 2008). In yeast, HR involves the RAD52 epistasis group (Krogh and Symington 2004) and uses a homologous sequence as a template for repair-typically the sister chromatid or, less frequently, the homologous chromosome (Kadyk and Hartwell 1992). Repair is initiated by 59-39 resection of the broken ends to form a 39 ssDNA overhang. This is a two-step process in which MRX/MRN (Mre11-Rad50-Xrs2 in Saccharomyces cerevisiae (Sung 1997;Sugawara et al. 2003;Wolner et al. 2003;Haruta et al. 2008). Stabilization of the nucleoprotein filament is achieved through interaction between Rad51 and Rad54; strand invasion is then initiated, resulting in the formation of a displacement (D) loop (Petukhova et al. 1998;Mazin et al. 2003;Sugawara et al. 2003). RPA further functions postsynaptically to stabilize DNA pairing and possibly the displaced ssD...

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Histone methyltransferase Dot1 and Rad9 inhibit single-stranded DNA accumulation at DSBs and uncapped telomeres

Cited by 147 publications

References 61 publications

A Genomewide Suppressor and Enhancer Analysis of cdc13-1 Reveals Varied Cellular Processes Influencing Telomere Capping in Saccharomyces cerevisiae

A Genomewide Suppressor and Enhancer Analysis of cdc13-1 Reveals Varied Cellular Processes Influencing Telomere Capping in Saccharomyces cerevisiae

TEN1Is Essential forCDC13-Mediated Telomere Capping

Break-induced ATR and Ddb1–Cul4^Cdt2 ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

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Histone methyltransferase Dot1 and Rad9 inhibit single-stranded DNA accumulation at DSBs and uncapped telomeres

Cited by 147 publications

References 61 publications

A Genomewide Suppressor and Enhancer Analysis of cdc13-1 Reveals Varied Cellular Processes Influencing Telomere Capping in Saccharomyces cerevisiae

A Genomewide Suppressor and Enhancer Analysis of cdc13-1 Reveals Varied Cellular Processes Influencing Telomere Capping in Saccharomyces cerevisiae

TEN1Is Essential forCDC13-Mediated Telomere Capping

Break-induced ATR and Ddb1–Cul4Cdt2 ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

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Product

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Break-induced ATR and Ddb1–Cul4^Cdt2 ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast