Mechanisms of dCMP transferase reactions catalyzed by mouse Rev1 protein.
The Rev1 protein, a member of a large family of translesion DNA polymerases, catalyzes a dCMP transfer reaction. Recombinant mouse Rev1 protein was found to insert a dCMP residue opposite guanine, adenine, thymine, cytosine, uracil, and an apurinic/apyrimidinic site and to have weak ability for transfer to a mismatched terminus. The mismatch-extension ability was strongly enhanced by a guanine residue on the template near the mismatched terminus; this was not the case with an apurinic/apyrimidinic site and the other template nucleotides. Kinetic analysis of the dCMP transferase reaction provided evidence for high affinity for dCTP with template G but not the other templates, whereas the template nucleotide did not much affect the V(max) value. Furthermore, it could be established that the mouse Rev1 protein inserts dGMP and dTMP residues opposite template guanine at a V(max) similar to that for dCMP.
Masuda
Yuji
Y
Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
Takahashi
Mamoru
M
Fukuda
Saburo
S
Sumii
Masaharu
M
Kamiya
Kenji
K
D000595:Amino Acid Sequence; D000818:Animals; D020137:Base Pair Mismatch; D001665:Binding Sites; D015152:Blotting, Northern; D002499:Centrifugation, Density Gradient; D004260:DNA Repair; D018076:DNA, Complementary / Q000378:metabolism; D003843:Deoxycytidine Monophosphate / Q000378:metabolism*; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism*; D006147:Guanine / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009154:Mutation; D009713:Nucleotidyltransferases; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D000099270:Y-Family DNA Polymerases
D018076:DNA, Complementary; D003843:Deoxycytidine Monophosphate; D003848:Deoxyguanine Nucleotides; D005656:Fungal Proteins; D006147:Guanine; D009713:Nucleotidyltransferases; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C007257:2'-deoxyguanosine 5'-phosphate; C100951:REV1 protein, S cerevisiae; D000099270:Y-Family DNA Polymerases