Loss of Dna2 fidelity results in decreased Exo1-mediated resection at DNA double-strand breaks
| dc.contributor.author | Mojumdar, Aditya | |
| dc.contributor.author | Granger, Courtney | |
| dc.contributor.author | Lunke, Martine | |
| dc.contributor.author | Cobb, Jennifer A. | |
| dc.date.accessioned | 2024-03-27T17:26:15Z | |
| dc.date.available | 2024-03-27T17:26:15Z | |
| dc.date.issued | 2024 | |
| dc.description | We thank Lorraine Symington for generously providing us with the plasmid overexpressing EXO1, pEM-EXO1. | |
| dc.description.abstract | A DNA double-strand break (DSB) is one of the most dangerous types of DNA damage that is repaired largely by homologous recombination or nonhomologous end-joining (NHEJ). The interplay of repair factors at the break directs which pathway is used, and a subset of these factors also function in more mutagenic alternative (alt) repair pathways. Resection is a key event in repair pathway choice and extensive resection, which is a hallmark of homologous recombination, and it is mediated by two nucleases, Exo1 and Dna2. We observed differences in resection and repair outcomes in cells harboring nuclease-dead dna2-1 compared with dna2Δ pif1-m2 that could be attributed to the level of Exo1 recovered at DSBs. Cells harboring dna2-1 showed reduced Exo1 localization, increased NHEJ, and a greater resection defect compared with cells where DNA2 was deleted. Both the resection defect and the increased rate of NHEJ in dna2-1 mutants were reversed upon deletion of KU70 or ectopic expression of Exo1. By contrast, when DNA2 was deleted, Exo1 and Ku70 recovery levels did not change; however, Nej1 increased as did the frequency of alt-end joining/microhomology-mediated end-joining repair. Our findings demonstrate that decreased Exo1 at DSBs contributed to the resection defect in cells expressing inactive Dna2 and highlight the complexity of understanding how functionally redundant factors are regulated in vivo to promote genome stability. | |
| dc.description.reviewstatus | Reviewed | |
| dc.description.scholarlevel | Faculty | |
| dc.description.sponsorship | This work was supported by operating grants from Canadian Institutes of Health Research (grant nos.: MOP-82736 and MOP-137062) and Natural Sciences and Engineering Research Council of Canada (grant no.: 418122) awarded to J. A. C. | |
| dc.identifier.citation | Mojumdar, A., Granger, C., Lunke, M., & Cobb, J. A. (2024). Loss of Dna2 fidelity results in decreased Exo1-mediated resection at DNA double-strand breaks, Journal of Biological Chemistry, 300(3), 105708. https://doi.org/10.1016/j.jbc.2024.105708 | |
| dc.identifier.uri | https://doi.org/10.1016/j.jbc.2024.105708 | |
| dc.identifier.uri | https://hdl.handle.net/1828/16302 | |
| dc.language.iso | en | |
| dc.publisher | Journal of Biological Chemistry | |
| dc.rights | Attribution 4.0 International | en |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
| dc.subject | DSB repair pathway choice | |
| dc.subject | Dna2 nuclease | |
| dc.subject | Exo1 nuclease | |
| dc.subject | mutant | |
| dc.subject | 5' DNA resection | |
| dc.subject | alternative end-joining | |
| dc.subject.department | Department of Microbiology and Biochemistry | |
| dc.subject.department | Department of Biochemistry and Microbiology | |
| dc.title | Loss of Dna2 fidelity results in decreased Exo1-mediated resection at DNA double-strand breaks | |
| dc.type | Article |