Co-localization in replication foci and interaction of human Y-family members, DNA polymerase pol eta and REVl protein.
The progress of replicative DNA polymerases along the replication fork may be impeded by the presence of lesions in the genome. One way to circumvent such hurdles involves the recruitment of specialized DNA polymerases that perform limited incorporation of nucleotides in the vicinity of the damaged site. This process entails DNA polymerase switch between replicative and specialized DNA polymerases. Five eukaryotic proteins can carry out translesion synthesis (TLS) of damaged DNA in vitro, DNA polymerases zeta, eta, iota, and kappa, and REV1. To identify novel proteins that interact with hpol eta, we performed a yeast two-hybrid screen. In this paper, we show that hREV1 interacts with hpol eta as well as with hpol kappa and poorly with hpol iota. Furthermore, cellular localization analysis demonstrates that hREV1 is present, with hpol eta in replication factories at stalled replication forks and is tightly associated with nuclear structures. This hREV1 nuclear localization occurs independently of the presence of hpol eta. Taken together, our data suggest a central role for hREV1 as a scaffold that recruits DNA polymerases involved in TLS.
Tissier
Agnès
A
UPR9OO3 - CNRS, Cancérogenèse et Mutagenèse Moléculaire et Structurale, ESBS, Pole API/Bd Sébastien Brant BP 10413, F-67412 - ILLKIRCH, France. tissier@esbs.u-strasbg.fr
Kannouche
Patricia
P
Reck
Marie-Pierre
MP
Lehmann
Alan R
AR
Fuchs
Robert P P
RP
Cordonnier
Agnès
A
D001483:Base Sequence; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016196:S Phase; D020798:Two-Hybrid System Techniques; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
D004247:DNA; D004259:DNA-Directed DNA Polymerase; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases; D011994:Recombinant Proteins; C429004:REV1 protein, human; D000099270:Y-Family DNA Polymerases; C112338:Rad30 protein