从黑腹果蝇中纯化的（6-4）光裂合酶对（6-4）光产物的非诱变修复

Non-mutagenic repair of (6-4)photoproducts by (6-4)photolyase purified from Drosophila melanogaster.

作者信息

Todo T, Ryo H, Borden A, Lawrence C, Sakaguchi K, Hirata H, Nomura T

机构信息

Department of Radiation Biology, Faculty of Medicine, Osaka Univ., Japan.

出版信息

Mutat Res. 1997 Nov;385(2):83-93. doi: 10.1016/s0921-8777(97)00045-1.

DOI:10.1016/s0921-8777(97)00045-1

PMID:9447230

Abstract

The (6-4)photoproduct DNA photolyase ((6-4)photolyase) repairs UV-induced pyrimidine (6-4) pyrimidone photoproduct ((6-4)photoproduct, pyr[6,4]pyr) in a light dependent manner. Drosophila (6-4)photolyase was purified to near homogeneity from Drosophila embryonic cells and is shown to be a 62 kDa protein as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified (6-4)photolyase repairs (6-4)photoproducts induced at 5'-CC-3' site (C[6,4]C) as well as T[6,4]T and T[6,4]C. Photoreactivation of (6-4)photoproduct constructed in M13 phage eliminates the replication block and abolishes induced mutagenesis in E. coli cells, suggesting that the (6-4)photolyase repairs the photoproduct to the unmodified form.

摘要

（6-4）光产物DNA光解酶（（6-4）光解酶）以光依赖的方式修复紫外线诱导的嘧啶（6-4）嘧啶酮光产物（（6-4）光产物，pyr[6,4]pyr）。从果蝇胚胎细胞中纯化出接近均一的果蝇（6-4）光解酶，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定其为62 kDa的蛋白质。纯化的（6-4）光解酶可修复在5'-CC-3'位点（C[6,4]C）以及T[6,4]T和T[6,4]C处诱导产生的（6-4）光产物。M13噬菌体中构建的（6-4）光产物的光复活消除了复制阻断并消除了大肠杆菌细胞中的诱导诱变，这表明（6-4）光解酶将光产物修复为未修饰的形式。