XRCC3 and Rad51 modulate replication fork progression on damaged vertebrate chromosomes.
The mechanisms by which the progression of eukaryotic replication forks is controlled after DNA damage are unclear. We have found that fork progression is slowed by cisplatin or UV treatment in intact vertebrate cells and in replication assays in vitro. Fork slowing is reduced or absent in irs1SF CHO cells and XRCC3(-/-) chicken DT40 cells, indicating that fork slowing is an active process that requires the homologous recombination protein XRCC3. The addition of purified human Rad51C-XRCC3 complex restores fork slowing in permeabilized XRCC3(-/-) cells. Moreover, the requirement for XRCC3 for fork slowing can be circumvented by addition of human Rad51. These data demonstrate that the recombination proteins XRCC3 and Rad51 cooperatively modulate the progression of replication forks on damaged vertebrate chromosomes.
Henry-Mowatt
Judith
J
School of Biological Sciences, University of Manchester, Stopford Building, Oxford Road, United Kingdom.
Jackson
Dean
D
Masson
Jean-Yves
JY
Johnson
Penny A
PA
Clements
Paula M
PM
Benson
Fiona E
FE
Thompson
Larry H
LH
Takeda
Shunichi
S
West
Stephen C
SC
Caldecott
Keith W
KW
D000818:Animals; D030161:Avian Proteins; D016466:CHO Cells; D002645:Chickens; D002875:Chromosomes / Q000235:genetics*; D002945:Cisplatin / Q000494:pharmacology; D006224:Cricetinae; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004261:DNA Replication / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D005057:Eukaryotic Cells / Q000378:metabolism*; D051135:Rad51 Recombinase; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D014466:Ultraviolet Rays
D030161:Avian Proteins; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C110901:X-ray repair cross complementing protein 3; C495150:RAD51 protein, Gallus gallus; D051135:Rad51 Recombinase; D002945:Cisplatin