The 9-1-1 Complex Controls Mre11 Nuclease and Checkpoint Activation during Short-Range Resection of DNA Double-Strand Breaks

Gobbini, Elisa; Casari, Erika; Colombo, Chiara Vittoria; Bonetti, Diego; Longhese, Maria Pia

doi:10.1016/j.celrep.2020.108287

Cited by 18 publications

(26 citation statements)

References 66 publications

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“…It is known that Mre11 is responsible for ∼300 bp of resection in exo1 Δ sgs1 Δ double mutant yeast defective in long range resection (35). Indeed, ddPCR quantification of ssDNA produced from resection in exo1 Δ sgs1 Δ yeast showed a clear decrease in resection efficiency in this strain as compared to WT (Figure 4A).…”

Section: Resultsmentioning

confidence: 99%

“…It is equally important to consider where resection intermediates were not observed, which was especially noticeable in exo1 sgs1 , a strain deficient in long range resection thought to mainly support cleavage by Mre11 (35). Resection initiation signal with this strain increased dramatically, although the signal at −65/−66 was only modestly increased relative to exo1 .…”

Section: Discussionmentioning

confidence: 99%

“…Consistently, the −206 signal was abrogated in exo1 yeast identifying it as a pause point during extension by Exo1. We do not know the nature of the obstacle that causes Exo1to pause but a strong candidate is the 9-1-1 complex based on recent observations (35). Whatever is responsible is again sequence specific as the location of the peak shifted with the DSB-proximal 3 bp deletions.…”

Section: Discussionmentioning

confidence: 99%

“…Consistently, the -206 signal was abrogated in exo1 yeast suggesting it as a pause point during extension by Exo1. We do not know the nature of the obstacle that causes Exo1 to pause but one candidate is the 9-1-1 complex based on recent observations (Gobbini et al, 2020).…”

Section: A Central Question Is What Determines the Pattern Of Cleavage At Mitotic Dsbs In Vivomentioning

confidence: 99%

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Mapping yeast mitotic 5’ resection at base resolution reveals the sequence and positional dependence of nucleasesin vivo

Bazzano

Lomonaco

Wilson

2021

Preprint

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Resection of the 5'-terminated strand at DNA double strand breaks (DSBs) is the critical regulated step in the transition to homologous recombination. Biochemical and genetic studies have led to a multi-step model of DSB resection in which endonucleolytic cleavage mediated by Mre11 in partnership with Sae2 is coupled with exonucleolytic cleavage mediated by redundant pathways catalyzed by Exo1 and Sgs1/Dna2. These models have not been well tested at mitotic DSBs in vivo because most methods commonly used to monitor resection cannot precisely map early cleavage events. Here we report resection monitoring with next-generation sequencing in which unique molecular identifiers allow exact counting of cleaved 5' ends at base pair resolution. Mutant strains, including exo1Δ, mre11-H125N, exo1Δ and exo1Δ sgs1Δ, revealed a major Mre11-dependent cleavage position 60 to 70 bp from the DSB end whose exact position depended on local sequence and tracked an apparent motif. They further revealed an Exo1-dependent pause point approximately 200 bp from the DSB. Suppressing resection extension in exo1Δ sgs1Δ yeast exposed a footprint of regions where cleavage was restricted within 119 bp of the DSB and near the Exo1 pause point and where it was much less restrained. These results provide detailed in vivo support of prevailing models of DSB resection and extend them to show the combined influence of sequence specificity and access restrictions on Mre11 and Exo1 nucleases.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: A Central Question Is What Determines the Pattern Of Cleavage At Mitotic Dsbs In Vivomentioning

confidence: 99%

See 2 more Smart Citations

Mapping yeast mitotic 5’ resection at base resolution reveals the sequence and positional dependence of nucleasesin vivo

Bazzano

Lomonaco

Wilson

2021

Preprint

View full text Add to dashboard Cite

show abstract

“…It is known that Mre11 is responsible for ∼300 bp of resection in exo1 Δ sgs1 Δ double mutant yeast defective in long range resection ( 38 ). Indeed, ddPCR quantification of ssDNA produced from resection in exo1 Δ sgs1 Δ yeast showed a decrease in resection efficiency in this strain as compared to WT (Figure 4A ).…”

Section: Resultsmentioning

confidence: 99%

Mapping yeast mitotic 5′ resection at base resolution reveals the sequence and positional dependence of nucleases in vivo

Bazzano

Lomonaco²,

Wilson

2021

Nucleic Acids Research

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Resection of the 5′-terminated strand at DNA double-strand breaks (DSBs) is the critical regulated step in the transition to homologous recombination. Recent studies have described a multi-step model of DSB resection where endonucleolytic cleavage mediated by Mre11 and Sae2 leads to further degradation mediated by redundant pathways catalyzed by Exo1 and Sgs1/Dna2. These models have not been well tested at mitotic DSBs in vivo because most methods used to monitor resection cannot precisely map early cleavage events. Here we report resection monitoring with high-throughput sequencing using molecular identifiers, allowing exact counting of cleaved 5′ ends at base resolution. Mutant strains, including exo1Δ, mre11-H125N and exo1Δ sgs1Δ, revealed a major Mre11-dependent cleavage position 60–70 bp from the DSB end whose exact position depended on local sequence. They further revealed an Exo1-dependent pause point approximately 200 bp from the DSB. Suppressing resection extension in exo1Δ sgs1Δ yeast exposed a footprint of regions where cleavage was restricted within 119 bp of the DSB. These results provide detailed in vivo views of prevailing models of DSB resection and extend them to show the combined influence of sequence specificity and access restrictions on Mre11 and Exo1 nucleases.

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Structures of 9-1-1 DNA checkpoint clamp loading at gaps from start to finish and ramification on biology

et al. 2023

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The 9-1-1 Complex Controls Mre11 Nuclease and Checkpoint Activation during Short-Range Resection of DNA Double-Strand Breaks

Cited by 18 publications

References 66 publications

Mapping yeast mitotic 5’ resection at base resolution reveals the sequence and positional dependence of nucleasesin vivo

Mapping yeast mitotic 5’ resection at base resolution reveals the sequence and positional dependence of nucleasesin vivo

Mapping yeast mitotic 5′ resection at base resolution reveals the sequence and positional dependence of nucleases in vivo

Structures of 9-1-1 DNA checkpoint clamp loading at gaps from start to finish and ramification on biology

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