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Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links
DNA interstrand cross-links (ICLs) are a form of DNA damage that requires the interplay of a number of repair proteins including those of the Fanconi anemia (FA) and the homologous recombination (HR) pathways. Pathogenic variants in the essential gene BRCA2/FANCD1, when monoallelic, predispose to br...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7263144/ https://www.ncbi.nlm.nih.gov/pubmed/32354836 http://dx.doi.org/10.1101/gad.336446.120 |
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author | Rickman, Kimberly A. Noonan, Raymond J. Lach, Francis P. Sridhar, Sunandini Wang, Anderson T. Abhyankar, Avinash Huang, Athena Kelly, Michael Auerbach, Arleen D. Smogorzewska, Agata |
author_facet | Rickman, Kimberly A. Noonan, Raymond J. Lach, Francis P. Sridhar, Sunandini Wang, Anderson T. Abhyankar, Avinash Huang, Athena Kelly, Michael Auerbach, Arleen D. Smogorzewska, Agata |
author_sort | Rickman, Kimberly A. |
collection | PubMed |
description | DNA interstrand cross-links (ICLs) are a form of DNA damage that requires the interplay of a number of repair proteins including those of the Fanconi anemia (FA) and the homologous recombination (HR) pathways. Pathogenic variants in the essential gene BRCA2/FANCD1, when monoallelic, predispose to breast and ovarian cancer, and when biallelic, result in a severe subtype of Fanconi anemia. BRCA2 function in the FA pathway is attributed to its role as a mediator of the RAD51 recombinase in HR repair of programmed DNA double-strand breaks (DSB). BRCA2 and RAD51 functions are also required to protect stalled replication forks from nucleolytic degradation during response to hydroxyurea (HU). While RAD51 has been shown to be necessary in the early steps of ICL repair to prevent aberrant nuclease resection, the role of BRCA2 in this process has not been described. Here, based on the analysis of BRCA2 DNA-binding domain (DBD) mutants (c.8488-1G>A and c.8524C>T) discovered in FA patients presenting with atypical FA-like phenotypes, we establish that BRCA2 is necessary for the protection of DNA at ICLs. Cells carrying BRCA2 DBD mutations are sensitive to ICL-inducing agents but resistant to HU treatment consistent with relatively high HR repair in these cells. BRCA2 function at an ICL protects against DNA2–WRN nuclease–helicase complex and not the MRE11 nuclease that is implicated in the resection of HU-induced stalled replication forks. Our results also indicate that unlike the processing at HU-induced stalled forks, the function of the SNF2 translocases (SMARCAL1, ZRANB3, or HLTF), implicated in fork reversal, are not an integral component of the ICL repair, pointing to a different mechanism of fork protection at different DNA lesions. |
format | Online Article Text |
id | pubmed-7263144 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-72631442020-12-01 Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links Rickman, Kimberly A. Noonan, Raymond J. Lach, Francis P. Sridhar, Sunandini Wang, Anderson T. Abhyankar, Avinash Huang, Athena Kelly, Michael Auerbach, Arleen D. Smogorzewska, Agata Genes Dev Research Paper DNA interstrand cross-links (ICLs) are a form of DNA damage that requires the interplay of a number of repair proteins including those of the Fanconi anemia (FA) and the homologous recombination (HR) pathways. Pathogenic variants in the essential gene BRCA2/FANCD1, when monoallelic, predispose to breast and ovarian cancer, and when biallelic, result in a severe subtype of Fanconi anemia. BRCA2 function in the FA pathway is attributed to its role as a mediator of the RAD51 recombinase in HR repair of programmed DNA double-strand breaks (DSB). BRCA2 and RAD51 functions are also required to protect stalled replication forks from nucleolytic degradation during response to hydroxyurea (HU). While RAD51 has been shown to be necessary in the early steps of ICL repair to prevent aberrant nuclease resection, the role of BRCA2 in this process has not been described. Here, based on the analysis of BRCA2 DNA-binding domain (DBD) mutants (c.8488-1G>A and c.8524C>T) discovered in FA patients presenting with atypical FA-like phenotypes, we establish that BRCA2 is necessary for the protection of DNA at ICLs. Cells carrying BRCA2 DBD mutations are sensitive to ICL-inducing agents but resistant to HU treatment consistent with relatively high HR repair in these cells. BRCA2 function at an ICL protects against DNA2–WRN nuclease–helicase complex and not the MRE11 nuclease that is implicated in the resection of HU-induced stalled replication forks. Our results also indicate that unlike the processing at HU-induced stalled forks, the function of the SNF2 translocases (SMARCAL1, ZRANB3, or HLTF), implicated in fork reversal, are not an integral component of the ICL repair, pointing to a different mechanism of fork protection at different DNA lesions. Cold Spring Harbor Laboratory Press 2020-06-01 /pmc/articles/PMC7263144/ /pubmed/32354836 http://dx.doi.org/10.1101/gad.336446.120 Text en © 2020 Rickman et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Research Paper Rickman, Kimberly A. Noonan, Raymond J. Lach, Francis P. Sridhar, Sunandini Wang, Anderson T. Abhyankar, Avinash Huang, Athena Kelly, Michael Auerbach, Arleen D. Smogorzewska, Agata Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links |
title | Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links |
title_full | Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links |
title_fullStr | Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links |
title_full_unstemmed | Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links |
title_short | Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand cross-links |
title_sort | distinct roles of brca2 in replication fork protection in response to hydroxyurea and dna interstrand cross-links |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7263144/ https://www.ncbi.nlm.nih.gov/pubmed/32354836 http://dx.doi.org/10.1101/gad.336446.120 |
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