Interactions in the error-prone postreplication repair proteins hREV1, hREV3, and hREV7.
Most mutations after DNA damage in yeast Saccharomyces cerevisiae are induced by error-prone translesion DNA synthesis employing scRev1 and DNA polymerase zeta that consists of scRev3 and scRev7 proteins. Recently, the human REV1 (hREV1) and REV3 (hREV3) genes were identified, and their products were revealed to be involved in UV-induced mutagenesis, as observed for their yeast counterparts. Human REV7 (hREV7) was also cloned, and its product was found to interact with hREV3, but the biological function of hREV7 remained unknown. We report here the analyses of precise interactions in the human REV proteins. The interaction between hREV1 and hREV7 was identified by the yeast two-hybrid library screening using a bait of hREV7, which was confirmed by in vitro and in vivo binding assays. The homodimerization of hREV7 was also detected in the two-hybrid analysis. In addition, the precise domains for interaction between hREV7 and hREV1 or hREV3 and for hREV7 homodimerization were determined. Although hREV7 interacts with both hREV1 and hREV3, a stable complex formation of the three proteins was undetectable in vitro. These findings suggest the possibility that hREV7 might play an important role in regulating the enzymatic activities of hREV1 and hREV3 for mutagenesis in response to DNA damage.
Murakumo
Y
Y
Department of Pathology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. murakumo@med.nagoya-u.ac.jp
Ogura
Y
Y
Ishii
H
H
Numata
S
S
Ichihara
M
M
Croce
C M
CM
Fishel
R
R
Takahashi
M
M
D002352:Carrier Proteins / Q000378:metabolism* / Q000502:physiology; D003001:Cloning, Molecular; D004249:DNA Damage / Q000502:physiology; D004260:DNA Repair; D004261:DNA Replication / Q000502:physiology; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism* / Q000502:physiology; D019281:Dimerization; D006367:HeLa Cells; D006801:Humans; D064246:Mad2 Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011506:Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D000099270:Y-Family DNA Polymerases
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D064246:Mad2 Proteins; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases; D011506:Proteins; C120552:MAD2L2 protein, human; C429004:REV1 protein, human; D000099270:Y-Family DNA Polymerases; C118285:REV3L protein, human