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首页> 外文期刊>Molecular and Cellular Biology >Identification of replication factor C from Saccharomyces cerevisiae: a component of the leading-strand DNA replication complex.
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Identification of replication factor C from Saccharomyces cerevisiae: a component of the leading-strand DNA replication complex.

机译：从酿酒酵母中鉴定复制因子C：前导链DNA复制复合物的组成部分。

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摘要

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.
机译：根据人类细胞在体外支持猿猴病毒40（SV40）DNA复制起源的DNA复制的能力，已从人细胞中分离出许多蛋白质。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δ共同起作用，以在复制叉处合成前导链。为了确定这些蛋白质在细胞染色体起源的DNA复制过程中是否发挥相似的作用，我们已开始鉴定来自酿酒酵母的功能同源蛋白质。来自酿酒酵母的RFC通过在酵母PCNA和RPA的存在下刺激底漆单链DNA模板上的酵母DNA聚合酶δ的能力来纯化。与人细胞类似，酿酒酵母的RFC（scRFC）具有相关的DNA激活的ATPase活性以及引物模板，结构特异性的DNA结合活性。通过类似于噬菌体T4和SV40 DNA体外复制系统，酵母RFC，PCNA，RPA和DNA聚合酶δ活性共同充当前导DNA复制复合体。现在，已经纯化了酿酒酵母的RFC，以前在酿酒酵母中已鉴定出了先前显示的体外SV40 DNA复制所需的所有七个细胞因子。

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