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PMID: 22987153 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

DNA2 and EXO1 in replication-coupled, homology-directed repair and in the interplay between HDR and the FA/BRCA network.

Cell cycle (Georgetown, Tex.) ·Vol. 11 ·No. 21 ·2012-11-01 ·Pages 3983-96

Karanja KK, Cox SW, Duxin JP, Stewart SA, Campbell JL

Abstract

During DNA replication, stalled replication forks and DSBs arise when the replication fork encounters ICLs (interstrand crosslinks), covalent protein/DNA intermediates or other discontinuities in the template. Recently, homologous recombination proteins have been shown to function in replication-coupled repair of ICLs in conjunction with the Fanconi anemia (FA) regulatory factors FANCD2-FANCI, and, conversely, the FA gene products have been shown to play roles in stalled replication fork rescue even in the absence of ICLs, suggesting a broader role for the FA network than previously appreciated. Here we show that DNA2 helicase/nuclease participates in resection during replication-coupled repair of ICLs and other replication fork stresses. DNA2 knockdowns are deficient in HDR (homology-directed repair) and the S phase checkpoint and exhibit genome instability and sensitivity to agents that cause replication stress. DNA2 is partially redundant with EXO1 in these roles. DNA2 interacts with FANCD2, and cisplatin induces FANCD2 ubiquitylation even in the absence of DNA2. DNA2 and EXO1 deficiency leads to ICL sensitivity but does not increase ICL sensitivity in the absence of FANCD2. This is the first demonstration of the redundancy of human resection nucleases in the HDR step in replication-coupled repair, and suggests that DNA2 may represent a new mediator of the interplay between HDR and the FA/BRCA pathway.

MeSH Terms
Antineoplastic Agents/toxicity Breast Neoplasms/metabolism,pathology Cell Line, Tumor Cisplatin/toxicity DNA Damage/drug effects DNA Helicases/antagonists & inhibitors,genetics,metabolism DNA Repair DNA Repair Enzymes/antagonists & inhibitors,genetics,metabolism Exodeoxyribonucleases/antagonists & inhibitors,genetics,metabolism Fanconi Anemia/metabolism,pathology Fanconi Anemia Complementation Group D2 Protein/metabolism Female Genomic Instability/drug effects HEK293 Cells Humans RNA Interference RNA, Small Interfering Ubiquitination
Chemicals
Antineoplastic Agents Fanconi Anemia Complementation Group D2 Protein RNA, Small Interfering EXO1 protein, human Exodeoxyribonucleases DNA Helicases DNA2 protein, human DNA Repair Enzymes Cisplatin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Karanja Kenneth K
Braun Laboratories, California Institute of Technology, Pasadena, CA, USA.
Cox Stephanie W
Duxin Julien P
Stewart Sheila A
Campbell Judith L
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Article Info
Journal
Cell cycle (Georgetown, Tex.)
Abbr.
Cell Cycle
ISSN
1551-4005
DOI
10.4161/cc.22215
Published
2012-11-01
Epub
2012-00-17
Pages
3983-96
Language
English
Region
United States
NLM ID
101137841
PMCID
PMC3507494
Subset
IM
Grants
NIGMS NIH HHS · GM100186 · United States
Corrections
CommentIn
PMID: 23099921
CommentIn
PMID: 23159855
External Links
PubMed source DOI: 10.4161/cc.22215 PMC Full Text View journal details
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