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Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection
The two major pathways of DNA double-strand break repair, nonhomologous end-joining and homologous recombination, are highly conserved from yeast to mammals. The regulation of 5′-DNA resection controls repair pathway choice and influences repair outcomes. Nej1 was first identified as a canonical NHE...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9117546/ https://www.ncbi.nlm.nih.gov/pubmed/35429499 http://dx.doi.org/10.1016/j.jbc.2022.101937 |
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author | Mojumdar, Aditya Adam, Nancy Cobb, Jennifer A. |
author_facet | Mojumdar, Aditya Adam, Nancy Cobb, Jennifer A. |
author_sort | Mojumdar, Aditya |
collection | PubMed |
description | The two major pathways of DNA double-strand break repair, nonhomologous end-joining and homologous recombination, are highly conserved from yeast to mammals. The regulation of 5′-DNA resection controls repair pathway choice and influences repair outcomes. Nej1 was first identified as a canonical NHEJ factor involved in stimulating the ligation of broken DNA ends, and more recently, it was shown to participate in DNA end-bridging and in the inhibition of 5′-resection mediated by the nuclease/helicase complex Dna2–Sgs1. Here, we show that Nej1 interacts with Sae2 to impact DSB repair in three ways. First, we show that Nej1 inhibits interaction of Sae2 with the Mre11–Rad50–Xrs2 complex and Sae2 localization to DSBs. Second, we found that Nej1 inhibits Sae2-dependent recruitment of Dna2 independently of Sgs1. Third, we determined that NEJ1 and SAE2 showed an epistatic relationship for end-bridging, an event that restrains broken DNA ends and reduces the frequency of genomic deletions from developing at the break site. Finally, we demonstrate that deletion of NEJ1 suppressed the synthetic lethality of sae2Δ sgs1Δ mutants, and that triple mutant viability was dependent on Dna2 nuclease activity. Taken together, these findings provide mechanistic insight to how Nej1 functionality inhibits the initiation of DNA resection, a role that is distinct from its involvement in end-joining repair at DSBs. |
format | Online Article Text |
id | pubmed-9117546 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-91175462022-05-21 Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection Mojumdar, Aditya Adam, Nancy Cobb, Jennifer A. J Biol Chem Research Article The two major pathways of DNA double-strand break repair, nonhomologous end-joining and homologous recombination, are highly conserved from yeast to mammals. The regulation of 5′-DNA resection controls repair pathway choice and influences repair outcomes. Nej1 was first identified as a canonical NHEJ factor involved in stimulating the ligation of broken DNA ends, and more recently, it was shown to participate in DNA end-bridging and in the inhibition of 5′-resection mediated by the nuclease/helicase complex Dna2–Sgs1. Here, we show that Nej1 interacts with Sae2 to impact DSB repair in three ways. First, we show that Nej1 inhibits interaction of Sae2 with the Mre11–Rad50–Xrs2 complex and Sae2 localization to DSBs. Second, we found that Nej1 inhibits Sae2-dependent recruitment of Dna2 independently of Sgs1. Third, we determined that NEJ1 and SAE2 showed an epistatic relationship for end-bridging, an event that restrains broken DNA ends and reduces the frequency of genomic deletions from developing at the break site. Finally, we demonstrate that deletion of NEJ1 suppressed the synthetic lethality of sae2Δ sgs1Δ mutants, and that triple mutant viability was dependent on Dna2 nuclease activity. Taken together, these findings provide mechanistic insight to how Nej1 functionality inhibits the initiation of DNA resection, a role that is distinct from its involvement in end-joining repair at DSBs. American Society for Biochemistry and Molecular Biology 2022-04-13 /pmc/articles/PMC9117546/ /pubmed/35429499 http://dx.doi.org/10.1016/j.jbc.2022.101937 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Mojumdar, Aditya Adam, Nancy Cobb, Jennifer A. Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection |
title | Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection |
title_full | Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection |
title_fullStr | Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection |
title_full_unstemmed | Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection |
title_short | Nej1 interacts with Sae2 at DNA double-stranded breaks to inhibit DNA resection |
title_sort | nej1 interacts with sae2 at dna double-stranded breaks to inhibit dna resection |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9117546/ https://www.ncbi.nlm.nih.gov/pubmed/35429499 http://dx.doi.org/10.1016/j.jbc.2022.101937 |
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