DNA2 and EXO1 in replication-coupled, homology-directed repair and in the interplay between HDR and the FA/BRCA network
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DNA2 and EXO1 in replication-coupled, homology-directed repair and in the interplay between HDR and the FA/BRCA network. / Karanja, Kenneth K; Cox, Stephanie W; Duxin, Julien P; Stewart, Sheila A; Campbell, Judith L.
In: Cell Cycle, Vol. 11, No. 21, 01.11.2012, p. 3983-96.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - DNA2 and EXO1 in replication-coupled, homology-directed repair and in the interplay between HDR and the FA/BRCA network
AU - Karanja, Kenneth K
AU - Cox, Stephanie W
AU - Duxin, Julien P
AU - Stewart, Sheila A
AU - Campbell, Judith L
PY - 2012/11/1
Y1 - 2012/11/1
N2 - 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.
AB - 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.
KW - Antineoplastic Agents
KW - Breast Neoplasms
KW - Cell Line, Tumor
KW - Cisplatin
KW - DNA Damage
KW - DNA Helicases
KW - DNA Repair
KW - DNA Repair Enzymes
KW - Exodeoxyribonucleases
KW - Fanconi Anemia
KW - Fanconi Anemia Complementation Group D2 Protein
KW - Female
KW - Genomic Instability
KW - HEK293 Cells
KW - Humans
KW - RNA Interference
KW - RNA, Small Interfering
KW - Ubiquitination
KW - Journal Article
KW - Research Support, N.I.H., Extramural
KW - Research Support, Non-U.S. Gov't
KW - Research Support, U.S. Gov't, Non-P.H.S.
U2 - 10.4161/cc.22215
DO - 10.4161/cc.22215
M3 - Journal article
C2 - 22987153
VL - 11
SP - 3983
EP - 3996
JO - Cell Cycle
JF - Cell Cycle
SN - 1538-4101
IS - 21
ER -
ID: 176967681