Mechanisms of dCMP Transferase Reactions Catalyzed by Mouse Rev1 Protein

• Published in: The Journal of biological chemistry Vol. 277; no. 4; pp. 3040 - 3046
• Main Authors: Masuda, Yuji, Takahashi, Mamoru, Fukuda, Saburo, Sumii, Masaharu, Kamiya, Kenji
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25.01.2002; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Animals; Base Pair Mismatch; Binding Sites; Blotting, Northern; Centrifugation, Density Gradient; Deoxycytidine Monophosphate - metabolism; Deoxyguanine Nucleotides - metabolism; DNA Repair; DNA, Complementary - metabolism; Electrophoresis, Polyacrylamide Gel; Fungal Proteins - metabolism; Guanine - chemistry; Humans; Kinetics; Mice; Mice, Inbred C57BL; Molecular Sequence Data; Mutation; Nucleotidyltransferases; Protein Binding; Protein Structure, Tertiary; Recombinant Proteins - metabolism; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; Tissue Distribution
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110149200

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 Vmax value. Furthermore, it could be established that the mouse Rev1 protein inserts dGMP and dTMP residues opposite template guanine at aVmax similar to that for dCMP.