Ubiquitin-binding motifs in REV1 protein are required for its role in the tolerance of DNA damage.
REV1 protein is a eukaryotic member of the Y family of DNA polymerases involved in the tolerance of DNA damage by replicative bypass. The precise role(s) of REV1 in this process is not known. Here we show, by using the yeast two-hybrid assay and the glutathione S-transferase pull-down assay, that mouse REV1 can physically interact with ubiquitin. The association of REV1 with ubiquitin requires the ubiquitin-binding motifs (UBMs) located at the C terminus of REV1. The UBMs also mediate the enhanced association between monoubiquitylated PCNA and REV1. In cells exposed to UV radiation, the association of REV1 with replication foci is dependent on functional UBMs. The UBMs of REV1 are shown to contribute to DNA damage tolerance and damage-induced mutagenesis in vivo.
Guo
Caixia
C
Laboratory of Molecular Pathology, Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9072, USA.
Tang
Tie-Shan
TS
Bienko
Marzena
M
Parker
Joanne L
JL
Bielen
Aleksandra B
AB
Sonoda
Eiichiro
E
Takeda
Shunichi
S
Ulrich
Helle D
HD
Dikic
Ivan
I
Friedberg
Errol C
EC
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002645:Chickens; D002522:Chlorocebus aethiops; D004249:DNA Damage; D004259:DNA-Directed DNA Polymerase; D005982:Glutathione Transferase / Q000378:metabolism; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin / Q000378:metabolism; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
D004259:DNA-Directed DNA Polymerase; D005982:Glutathione Transferase; D009713:Nucleotidyltransferases; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; C496086:Rev1 protein, mouse; D000099270:Y-Family DNA Polymerases