REV1 protein interacts with PCNA: significance of the REV1 BRCT domain in vitro and in vivo.
REV1 protein, a eukaryotic member of the Y family of DNA polymerases, is involved in the tolerance of DNA damage by translesion DNA synthesis. It is unclear how REV1 is recruited to replication foci in cells. Here, we report that mouse REV1 can bind directly to PCNA and that monoubiquitylation of PCNA enhances this interaction. The interaction between REV1 protein and PCNA requires a functional BRCT domain located near the N terminus of the former protein. Deletion or mutational inactivation of the BRCT domain abolishes the targeting of REV1 to replication foci in unirradiated cells, but not in UV-irradiated cells. In vivo studies in both chicken DT40 cells and yeast directly support the requirement of the BRCT domain of REV1 for cell survival and DNA damage-induced mutagenesis.
Guo
Caixia
C
Laboratory of Molecular Pathology, Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
Sonoda
Eiichiro
E
Tang
Tie-Shan
TS
Parker
Joanne L
JL
Bielen
Aleksandra B
AB
Takeda
Shunichi
S
Ulrich
Helle D
HD
Friedberg
Errol C
EC
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D002645:Chickens; D004249:DNA Damage; D018076:DNA, Complementary / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000502:physiology; D004259:DNA-Directed DNA Polymerase; D051379:Mice; D016296:Mutagenesis; D009713:Nucleotidyltransferases / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D000099270:Y-Family DNA Polymerases
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D009713:Nucleotidyltransferases; D018809:Proliferating Cell Nuclear Antigen; D025801:Ubiquitin; C496086:Rev1 protein, mouse; D000099270:Y-Family DNA Polymerases