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A role for human homologous recombination factors in suppressing microhomology-mediated end joining

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Nucl. Acids Res.-2016-Ahrabi-5743-57.pdfPublished version7.35 MBAdobe PDFView/Open
Title: A role for human homologous recombination factors in suppressing microhomology-mediated end joining
Authors: Ahrabi, S
Sarkar, S
Pfister, SX
Pirovano, G
Higgins, GS
Porter, AC
Humphrey, TC
Item Type: Journal Article
Abstract: DNA double-strand breaks (DSBs) are toxic lesions, which if improperly repaired can result in cell death or genomic instability. DSB repair is usually facilitated by the classical non-homologous end joining (C-NHEJ), or homologous recombination (HR) pathways. However, a mutagenic alternative NHEJ pathway, microhomology-mediated end joining (MMEJ), can also be deployed. While MMEJ is suppressed by C-NHEJ, the relationship between HR and MMEJ is less clear. Here, we describe a role for HR genes in suppressing MMEJ in human cells. By monitoring DSB mis-repair using a sensitive HPRT assay, we found that depletion of HR proteins, including BRCA2, BRCA1 or RPA, resulted in a distinct mutational signature associated with significant increases in break-induced mutation frequencies, deletion lengths and the annealing of short regions of microhomology (2-6 bp) across the break-site. This signature was dependent on CtIP, MRE11, POLQ and PARP, and thus indicative of MMEJ. In contrast to CtIP or MRE11, depletion of BRCA1 resulted in increased partial resection and MMEJ, thus revealing a functional distinction between these early acting HR factors. Together these findings indicate that HR factors suppress mutagenic MMEJ following DSB resection.
Issue Date: 8-Jul-2016
Date of Acceptance: 14-Apr-2016
URI: http://hdl.handle.net/10044/1/34553
DOI: 10.1093/nar/gkw326
ISSN: 1362-4962
Publisher: Oxford University Press (OUP)
Start Page: 5743
End Page: 5757
Journal / Book Title: Nucleic Acids Research
Volume: 44
Issue: 12
Copyright Statement: © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: BB/H003371/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
DOUBLE-STRAND BREAKS
DNA-POLYMERASE-THETA
LIGASE-III
REPAIR PATHWAY
DIRECTED REPAIR
BRCA1
DAMAGE
CTIP
RESECTION
FIDELITY
BRCA1 Protein
BRCA2 Protein
Base Sequence
Biological Assay
Carrier Proteins
Cell Line, Tumor
DNA
DNA Breaks, Double-Stranded
DNA End-Joining Repair
DNA-Binding Proteins
DNA-Directed DNA Polymerase
Endodeoxyribonucleases
Epithelial Cells
Fibroblasts
Humans
MRE11 Homologue Protein
Mutation
Nuclear Proteins
Osteoblasts
Poly(ADP-ribose) Polymerases
RNA, Small Interfering
Recombinational DNA Repair
Replication Protein A
Sequence Alignment
Sequence Homology, Nucleic Acid
Cell Line, Tumor
Fibroblasts
Osteoblasts
Epithelial Cells
Humans
Poly(ADP-ribose) Polymerases
DNA-Directed DNA Polymerase
Carrier Proteins
DNA-Binding Proteins
BRCA1 Protein
BRCA2 Protein
Nuclear Proteins
RNA, Small Interfering
DNA
Biological Assay
Sequence Alignment
Base Sequence
Sequence Homology, Nucleic Acid
Mutation
Replication Protein A
DNA Breaks, Double-Stranded
DNA End-Joining Repair
Recombinational DNA Repair
MRE11 Homologue Protein
05 Environmental Sciences
06 Biological Sciences
08 Information and Computing Sciences
Developmental Biology
Publication Status: Published
Online Publication Date: 2016-04-29
Appears in Collections:Department of Immunology and Inflammation
Faculty of Medicine