IDENTIFICATION OF REPLICATION FACTOR-C FROM SACCHAROMYCES-CEREVISIAE - A COMPONENT OF THE LEADING-STRAND DNA-REPLICATION COMPLEX
IDENTIFICATION OF REPLICATION FACTOR-C FROM SACCHAROMYCES-CEREVISIAE - A COMPONENT OF THE LEADING-STRAND DNA-REPLICATION COMPLEX
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DOI：
10.1128/mcb.12.1.155
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发表时间：
1992-01-01
影响因子：
5.3
通讯作者：
STILLMAN, B
中科院分区：
文献类型：
--
作者：
FIEN, K;STILLMAN, B
从人类细胞中分离出一些蛋白质，基于它们支持体外DNA复制的能力，起源于猴病毒40(SV40)的DNA复制。一种这样的蛋白质，复制因子C(RFC)，与增殖细胞核抗原(PCNA)、复制蛋白A(RPA)和DNA聚合酶Delta一起作用于复制叉处合成前导链。为了确定这些蛋白质是否在从细胞染色体中复制DNA的过程中发挥类似的作用，我们已经开始鉴定来自酿酒酵母的功能同源蛋白质。酿酒酵母RFC在酵母增殖细胞核抗原和RPA存在的情况下，通过在单链DNA模板上刺激酵母DNA聚合酶Delta的能力而得到纯化。与人类细胞一样，酿酒酵母RFC(ScRFC)具有DNA激活的ATPase活性以及引物-模板、结构特异性DNA结合活性。通过与噬菌体T4和SV40体外DNA复制系统的类似，酵母RFC、增殖细胞核抗原、RPA和DNA聚合酶Delta活性共同发挥着DNA复制复合体的主导作用。现在，酿酒酵母的RFC已经被提纯，以前被证明在体外复制SV40DNA所需的所有七个细胞因子都在酿酒酵母中被鉴定出来。
A number of proteins have been isolated from human cells on the basis of their ability to support DNA replication in vitro of the simian virus 40 (SV40) origin of DNA replication. One such protein, replication factor C (RFC), functions with the proliferating cell nuclear antigen (PCNA), replication protein A (RPA), and DNA polymerase delta to synthesize the leading strand at a replication fork. To determine whether these proteins perform similar roles during replication of DNA from origins in cellular chromosomes, we have begun to characterize functionally homologous proteins from the yeast Saccharomyces cerevisiae. RFC from S. cerevisiae was purified by its ability to stimulate yeast DNA polymerase delta on a primed single-stranded DNA template in the presence of yeast PCNA and RPA. Like its human-cell counterpart, RFC from S. cerevisiae (scRFC) has an associated DNA-activated ATPase activity as well as a primer-template, structure-specific DNA binding activity. By analogy with the phage T4 and SV40 DNA replication in vitro systems, the yeast RFC, PCNA, RPA, and DNA polymerase delta activities function together as a leading-strand DNA replication complex. Now that RFC from S. cerevisiae has been purified, all seven cellular factors previously shown to be required for SV40 DNA replication in vitro have been identified in S. cerevisiae.