Deoxycytidyl transferase activity of the human REV1 protein is closely associated with the conserved polymerase domain
Deoxycytidyl transferase activity of the human REV1 protein is closely associated with the conserved polymerase domain
复制标题
DOI：
10.1074/jbc.m008082200
复制
发表时间：
2001-05-04
影响因子：
4.8
通讯作者：
Kamiya, K
中科院分区：
文献类型：
--
作者：
Masuda, Y;Takahashi, M;Kamiya, K
REV 1蛋白是不断增长的跨损伤DNA聚合酶家族的成员。我们最初分离的人REV 1基因的cDNA编码1250个氨基酸残基，比以前报道的短一个氨基酸。REV 1的缩写形式被命名为REV 1 S。所有受检个体均表达等量的REV 1 S和REV 1 mRNA，表明REV 1 S mRNA是一种剪接变体。我们表明，REV 1 S蛋白也具有脱氧胞苷酰转移酶活性，插入一个dCMP对DNA模板脱嘌呤/脱嘧啶位点。REV 1 S蛋白的缺失和点突变分析显示，REV 1 S蛋白的脱氧胞苷酰转移酶和DNA结合活性所需的结构域位于translesion DNA聚合酶的保守结构域中。这一结果表明，脱氧胞苷酰转移酶的催化位点的结构非常相似的translesion DNA聚合酶。因此，REV 1 S蛋白以及可能还有REV 1蛋白的dCMP转移反应的分子机制可能与跨损伤DNA聚合酶的dNTP转移反应的分子机制相同。
The REV1 protein is a member of the growing family of translesion DNA polymerases. A cDNA of the human REV1 gene that we had originally isolated encoded 1250 amino acids residues, which was one amino acid shorter than previously reported ones. The shorter form of REV1 was named REV1S. All individuals examined expressed equivalent amounts of REV1S and REV1 mRNA, suggesting that the REV1S mRNA is a splicing variant. We show that the REV1S protein also possesses deoxycytidyl transferase activity that inserts a dCMP opposite a DNA template apurinic/apyrimidinic site. Deletion and point mutation analysis of the REV1S protein revealed that the domain required for deoxycytidyl transferase and DNA binding activities of the REV1S protein are located in a conserved domain of translesion DNA polymerases. This result indicates that the structure of the catalytic site of the deoxycytidyl transferase closely resembles that of the translesion DNA polymerases. Therefore, the molecular mechanism of the dCMP transfer reaction of the REV1S protein and maybe also the REV1 protein might be the same as that of the dNTP transfer reaction of the translesion DNA polymerases.