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一种必需的酿酒酵母单链DNA结合蛋白与人类RP-A的大亚基同源。

An essential Saccharomyces cerevisiae single-stranded DNA binding protein is homologous to the large subunit of human RP-A.

作者信息

Heyer W D, Rao M R, Erdile L F, Kelly T J, Kolodner R D

机构信息

Laboratory of Molecular Genetics, Dana-Farber Cancer Institute, Boston, MA 02115.

出版信息

EMBO J. 1990 Jul;9(7):2321-9. doi: 10.1002/j.1460-2075.1990.tb07404.x.

DOI:10.1002/j.1460-2075.1990.tb07404.x
PMID:2192864
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC551959/
Abstract

Single-stranded DNA binding proteins (SSBs) are known to play a role in DNA replication and recombination in prokaryotes. An SSB was previously purified from the yeast Saccharomyces cerevisiae. This SSB stimulated the activity of a cognate strand exchange protein (SEP1) in vitro suggesting a role in recombination. We have cloned and functionally analyzed the gene encoding this protein. DNA sequencing of the cloned DNA revealed a 621 amino acid open reading frame with a coding potential for a Mr 70,269 polypeptide. Highly significant amino acid homology was detected between this S.cerevisiae gene and the Mr 70,000 subunit polypeptide of human RP-A, a cellular protein essential for SV40 DNA replication in vitro. Therefore, we named the S.cerevisiae gene RPA1. RPA1 encodes an essential function in this organism as shown by tetrad analysis of heterozygous insertion mutants and is continuously required for mitotic growth. Cells lacking RPA1 accumulate as multiply budded cells with a single nucleus suggesting a defect in DNA replication.

摘要

已知单链DNA结合蛋白（SSB）在原核生物的DNA复制和重组中发挥作用。此前已从酿酒酵母中纯化出一种SSB。这种SSB在体外刺激了同源链交换蛋白（SEP1）的活性，表明其在重组中发挥作用。我们已经克隆并对编码该蛋白的基因进行了功能分析。对克隆DNA的测序揭示了一个621个氨基酸的开放阅读框，其编码潜力为一个70269道尔顿的多肽。在这个酿酒酵母基因与人类RP-A的70000道尔顿亚基多肽之间检测到高度显著的氨基酸同源性，RP-A是一种体外SV40 DNA复制所必需的细胞蛋白。因此，我们将酿酒酵母基因命名为RPA1。如杂合插入突变体的四分体分析所示，RPA1在该生物体中编码一种基本功能，并且有丝分裂生长持续需要它。缺乏RPA1的细胞会积累成具有单核的多芽细胞，这表明DNA复制存在缺陷。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/19d3281f780c/emboj00234-0289-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/6aa357e4e973/emboj00234-0287-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/3362b69e58a0/emboj00234-0288-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/19d3281f780c/emboj00234-0289-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/6aa357e4e973/emboj00234-0287-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/3362b69e58a0/emboj00234-0288-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/84c4/551959/19d3281f780c/emboj00234-0289-a.jpg

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