AtREV1, a Y-family DNA polymerase in Arabidopsis, has deoxynucleotidyl transferase activity

Takahashi, Shinya*; Sakamoto, Ayako; Tanaka, Atsushi; Shimizu, Kikuo*

To clarify the functions of AtREV1 protein, we expressed it in E. coli and purified it. The deoxynucleotidyl transferase activity of the recombinant AtREV1 was examined in a primer extension assay . The recombinant AtREV1 transferred one or two nucleotides to the primer end. Especially, it efficiently inserted dCMP regardless of the opposite base. AtREV1 also inserted a dCMP opposite the apurinic/apyrimidinic (AP) sites, which are physiologically generated or induced by various DNA-damaging agents. However, AtREV1 had no insertion activities against UV-inducible DNA lesions. Although the substrate specificity of AtREV1 was rather narrow in the presence of magnesium ion, it widened in the presence of manganese ion. These results suggest that AtREV1 serves as a deoxycytidyl transferase in plant cells.