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显性负性大鼠DNA聚合酶β突变体干扰酿酒酵母中的碱基切除修复。

Dominant negative rat DNA polymerase beta mutants interfere with base excision repair in Saccharomyces cerevisiae.

作者信息

Clairmont C A, Sweasy J B

机构信息

Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

出版信息

J Bacteriol. 1996 Feb;178(3):656-61. doi: 10.1128/jb.178.3.656-661.1996.

DOI:10.1128/jb.178.3.656-661.1996
PMID:8550496
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC177708/
Abstract

DNA polymerase beta is one of the smallest known eukaryotic DNA polymerases. This polymerase has been very well characterized in vitro, but its functional role in vivo has yet to be determined. Using a novel competition assay in Escherichia coli, we isolated two DNA polymerase beta dominant negative mutants. When we overexpressed the dominant negative mutant proteins in Saccharomyces cerevisiae, the cells became sensitive to methyl methanesulfonate. Interestingly, overexpression of the same polymerase beta mutant proteins did not confer sensitivity to UV damage, strongly suggesting that the mutant proteins interfere with the process of base excision repair but not nucleotide excision repair in S. cerevisiae. Our data implicate a role for polymerase IV, the S. cerevisiae polymerase beta homolog, in base excision repair in S. cerevisiae.

摘要

DNA聚合酶β是已知最小的真核生物DNA聚合酶之一。这种聚合酶在体外已得到很好的表征，但其在体内的功能作用尚未确定。利用大肠杆菌中的一种新型竞争测定法，我们分离出了两个DNA聚合酶β显性负突变体。当我们在酿酒酵母中过表达显性负突变蛋白时，细胞对甲磺酸甲酯变得敏感。有趣的是，相同的聚合酶β突变蛋白的过表达并未赋予对紫外线损伤的敏感性，这强烈表明突变蛋白干扰了酿酒酵母中的碱基切除修复过程，而不是核苷酸切除修复过程。我们的数据表明酿酒酵母中与DNA聚合酶β同源的聚合酶IV在碱基切除修复中发挥作用。

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本文引用的文献

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Acetylornithinase of Escherichia coli: partial purification and some properties.大肠杆菌的乙酰鸟氨酸酶：部分纯化及某些性质
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Detection and characterization of mammalian DNA polymerase beta mutants by functional complementation in Escherichia coli.通过在大肠杆菌中进行功能互补来检测和鉴定哺乳动物DNA聚合酶β突变体
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DNA repair synthesis during base excision repair in vitro is catalyzed by DNA polymerase epsilon and is influenced by DNA polymerases alpha and delta in Saccharomyces cerevisiae.在酿酒酵母中，体外碱基切除修复过程中的DNA修复合成由DNA聚合酶ε催化，并受DNA聚合酶α和δ的影响。
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Yeast open reading frame YCR14C encodes a DNA beta-polymerase-like enzyme.酵母开放阅读框YCR14C编码一种DNAβ聚合酶样酶。
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6
Purification and characterization of a new DNA polymerase from budding yeast Saccharomyces cerevisiae. A probable homolog of mammalian DNA polymerase beta.来自芽殖酵母酿酒酵母的一种新型DNA聚合酶的纯化与特性分析。一种可能与哺乳动物DNA聚合酶β同源的蛋白。
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DNA polymerase delta is required for base excision repair of DNA methylation damage in Saccharomyces cerevisiae.酿酒酵母中DNA甲基化损伤的碱基切除修复需要DNA聚合酶δ。
Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9047-51. doi: 10.1073/pnas.91.19.9047.
8
The yeast Saccharomyces cerevisiae DNA polymerase IV: possible involvement in double strand break DNA repair.酿酒酵母DNA聚合酶IV：可能参与双链断裂DNA修复。
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DNA polymerase beta conducts the gap-filling step in uracil-initiated base excision repair in a bovine testis nuclear extract.在牛睾丸核提取物中，DNA聚合酶β在尿嘧啶引发的碱基切除修复过程中执行填补缺口步骤。
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