一种与SV40 DNA复制核心起点结合的细胞蛋白的部分纯化及特性分析

Partial purification and characterization of a cellular protein that binds to the SV40 core origin of DNA replication.

作者信息

Alliger P, Traut W, Carstens E, Fanning E

机构信息

Institut für Biochemie, München, F.R.G.

出版信息

Biochim Biophys Acta. 1988 Dec 20;951(2-3):396-402. doi: 10.1016/0167-4781(88)90112-1.

DOI:10.1016/0167-4781(88)90112-1

PMID:2850016

Abstract

A monkey cell factor that interacts specifically with double- and single-stranded DNA sequences in the early domain of the simian virus 40 (SV40) core origin of replication was identified using gel-retention assays. The protein was enriched over 1200-fold using ion-exchange and affinity chromatography on single-strand DNA cellulose. Binding of protein to mutant origin DNA restriction fragments was correlated with replication activity of the mutant DNAs. Exonuclease footprint experiments on single-stranded DNA revealed prominent pause sites in the early domain of the core origin. The results suggest that this cellular protein may be involved in SV40 DNA replication.

摘要

利用凝胶滞留试验鉴定出一种猴细胞因子，它能与猿猴病毒40（SV40）核心复制起点早期区域的双链和单链DNA序列特异性相互作用。通过离子交换和在单链DNA纤维素上的亲和层析，该蛋白质被富集了1200多倍。蛋白质与突变型起点DNA限制片段的结合与突变型DNA的复制活性相关。对单链DNA进行的核酸外切酶足迹实验揭示了核心起点早期区域存在显著的停顿位点。结果表明，这种细胞蛋白可能参与SV40 DNA复制。