Nej1 Interacts with Mre11 to Regulate Tethering and Dna2 Binding at DNA Double-Strand Breaks

Mojumdar, Aditya; Sorenson, Kyle; Hohl, Marcel; Toulouze, Mathias; Lees‐Miller, Susan P.; Dubrana, Karine; Petrini, John H.J.; Cobb, Jennifer A.

doi:10.1016/j.celrep.2019.07.018

Cited by 30 publications

(124 citation statements)

References 43 publications

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“…Nej1 acetyl-mimicking mutant shows reduced binding to Lif1 and Yku80 in vitro, which correlates with increased resection seen in vivo (Fig 5D-F). It would be interesting to determine how these individual lysines affect interaction of Nej1 with other NHEJ proteins and whether the tethering activity of Nej1 itself is impaired in these mutants (56). In both yeast and mammals, it has been documented that Ku may interact with various histone deacetylases Our data supports a model (Fig 6) in which NuA4 is recruited by the MRX complex in the early steps after DSB formation.…”

Section: Discussionsupporting

confidence: 79%

Antagonistic relationship of NuA4 with the Non-Homologous End-Joining machinery at DNA damage sites

Ahmad

Côté

Cheng

et al. 2021

Preprint

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The NuA4 histone acetyltransferase complex, apart from its known role in gene regulation, has also been directly implicated in the repair of DNA double-strand breaks (DSBs), favoring homologous recombination (HR) in S/G2 during the cell cycle. Here, we investigate the antagonistic relationship of NuA4 with non-homologous end joining (NHEJ) factors. We detect NuA4 at DSBs in G1 when genes for NHEJ factors Rad9, Yku80 and Nej1 are mutated. This is accompanied with the binding of single-strand DNA binding protein RPA, indicating DNA end resection in G1. This NuA4 recruitment to DSBs depends on both Xrs2 and Lcd1/Ddc2. Introducing an acetyltransferase defective allele in these NHEJ mutant backgrounds decreases their hyper-resection phenotype in G1. Interestingly, we identified two novel non-histone acetylation targets of NuA4, Nej1 and Yku80. Acetyl-mimicking mutant of Nej1 inhibits NHEJ, decreases its interaction with other NHEJ factors and favors resection. Altogether, these results establish an antagonistic regulatory function of NuA4 and NHEJ factors in repair pathway choice and suggest a role of NuA4 in alternative repair mechanism that involves DNA-end resection in G1.

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Section: Discussionsupporting

confidence: 79%

Antagonistic relationship of NuA4 with the Non-Homologous End-Joining machinery at DNA damage sites

Ahmad

Côté

Cheng

et al. 2021

Preprint

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“…We asked whether the partial CPT sensitivity and sporulation defect observed for rad50-L1240F might be due to an effect on Mre11 nuclease activity. Using a Q-PCR based assay for resection of an HO DSB [29], we noted reduced DSB resection in rad50-L1240F and to a lesser extent rad50-D67Y (Fig 2F). However, Mre11 nuclease activity is minimally affected, since in contrast to mre11 nuclease dead alleles (mre11-3, mre11-H125N) [30,31], rad50-L1240F, rad50-D67Y, and mre11-E38K exhibit only mild synergy with exo1Δ or sgs1Δ with respect to CPT or MMS-sensitivity (S1B Fig).…”

Section: Plos Geneticsmentioning

confidence: 94%

“…RsaI cut DNA was normalized to uncut DNA as previously described to quantify the percentage of ssDNA per total amount of DNA [52]. Same primers were used for Q-PCR as previously published [29].…”

Section: Q-pcr Based Resection Assaymentioning

confidence: 99%

Modeling cancer genomic data in yeast reveals selection against ATM function during tumorigenesis

et al. 2020

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The DNA damage response (DDR) comprises multiple functions that collectively preserve genomic integrity and suppress tumorigenesis. The Mre11 complex and ATM govern a major axis of the DDR and several lines of evidence implicate that axis in tumor suppression. Components of the Mre11 complex are mutated in approximately five percent of human cancers. Inherited mutations of complex members cause severe chromosome instability syndromes, such as Nijmegen Breakage Syndrome, which is associated with strong predisposition to malignancy. And in mice, Mre11 complex mutations are markedly more susceptible to oncogene-induced carcinogenesis. The complex is integral to all modes of DNA double strand break (DSB) repair and is required for the activation of ATM to effect DNA damage signaling. To understand which functions of the Mre11 complex are important for tumor suppression, we undertook mining of cancer genomic data from the clinical sequencing program at Memorial Sloan Kettering Cancer Center, which includes the Mre11 complex among the 468 genes assessed. Twenty five mutations in MRE11 and RAD50 were modeled in S. cerevisiae and in vitro. The mutations were chosen based on recurrence and conservation between human and yeast. We found that a significant fraction of tumorborne RAD50 and MRE11 mutations exhibited separation of function phenotypes wherein Tel1/ATM activation was severely impaired while DNA repair functions were mildly or not affected. At the molecular level, the gene products of RAD50 mutations exhibited defects in ATP binding and hydrolysis. The data reflect the importance of Rad50 ATPase activity for Tel1/ATM activation and suggest that inactivation of ATM signaling confers an advantage to burgeoning tumor cells.

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“…Likewise, γH2Ax foci are not contiguous when visualized by highresolution microscopy, which revealed spatially distinct nanodomains clustered around DSB sites 18 . The mechanisms that sculpt γH2Ax domains have important implications, especially given the critical role of these platforms in damage responses, including: (1) tethering broken chromosomes until they are repaired 11,19 (2) repression of transcription 20,21 , and (3) sequestration of DDR factors around a DSB site 18,22 .…”

mentioning

confidence: 99%

DNA double-strand breaks induce H2Ax phosphorylation domains in a contact-dependent manner

et al. 2020

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Efficient repair of DNA double-strand breaks (DSBs) requires a coordinated DNA Damage Response (DDR), which includes phosphorylation of histone H2Ax, forming γH2Ax. This histone modification spreads beyond the DSB into neighboring chromatin, generating a DDR platform that protects against end disassociation and degradation, minimizing chromosomal rearrangements. However, mechanisms that determine the breadth and intensity of γH2Ax domains remain unclear. Here, we show that chromosomal contacts of a DSB site are the primary determinants for γH2Ax landscapes. DSBs that disrupt a topological border permit extension of γH2Ax domains into both adjacent compartments. In contrast, DSBs near a border produce highly asymmetric DDR platforms, with γH2Ax nearly absent from one broken end. Collectively, our findings lend insights into a basic DNA repair mechanism and how the precise location of a DSB may influence genome integrity.

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Nej1 Interacts with Mre11 to Regulate Tethering and Dna2 Binding at DNA Double-Strand Breaks

Abstract: Highlights d The MRX complex stabilizes Nej1 at the break site d Nej1 inhibits Dna2 recruitment and HR d Nej1 and the coiled-coil region of Rad50 are important for tethering broken DNA ends d Defects in 5 0 resection and end-tethering lead to large chromosome deletions at the DSB

Cited by 30 publications

References 43 publications

Antagonistic relationship of NuA4 with the Non-Homologous End-Joining machinery at DNA damage sites

Antagonistic relationship of NuA4 with the Non-Homologous End-Joining machinery at DNA damage sites

Modeling cancer genomic data in yeast reveals selection against ATM function during tumorigenesis

DNA double-strand breaks induce H2Ax phosphorylation domains in a contact-dependent manner

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