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大肠杆菌中缺乏DNA聚合酶I 5'→3'核酸外切酶活性和核酸外切酶VII的突变体中的脱氧核糖核酸修复

Deoxyribonucleic acid repair in Escherichia coli mutants deficient in the 5'----3' exonuclease activity of deoxyribonucleic acid polymerase I and exonuclease VII.

作者信息

Chase J W, Masker W E

出版信息

J Bacteriol. 1977 May;130(2):667-75. doi: 10.1128/jb.130.2.667-675.1977.

Abstract

A series of Escherichia coli strains deficient in the 5'----3' exonuclease activity associated with deoxyribonucleic acid (DNA) polymerase I (exonuclease VI) and exonuclease VII has been constructed. Both of these enzymes are capable of pyrimidine dimer excision in vitro. These strains were examined for conditional lethality, sensitivity to ultraviolet (UV) and X-irradiation, postirradiation DNA degradation, and ability to excise pyrimidine dimers. It was found that strains deficient in both exonuclease VI (polAex-) and exonuclease VII (xseA-) are significantly reduced in their ability to survive incubation at elevated temperature (43 degrees C) beyond the reduction previously observed for the polAex single mutants. The UV and X-ray sensitivity of the exonuclease VI-deficient strains was not increased by the addition of the xseA7 mutation. Mutants deficient in both enzymes are about as efficient as wild-type strains at excising dimers produced by up to 40 J/m2 UV. At higher doses strains containing only polAex- mutations show reduced ability to excise dimers; however, the interpretation of dimer excision data at these doses is complicated by extreme postirradiation DNA degradation in these strains. The additional deficiency in the polAex xseA7 double-mutant strains has no significant effect on either postirradiation DNA degradation or the apparent deficiency in dimer excision at high UV doses observed in polAex single mutants.

摘要

构建了一系列与脱氧核糖核酸(DNA)聚合酶I(核酸外切酶VI)和核酸外切酶VII相关的5'→3'核酸外切酶活性缺陷的大肠杆菌菌株。这两种酶在体外都能够切除嘧啶二聚体。对这些菌株进行了条件致死性、对紫外线(UV)和X射线照射的敏感性、照射后DNA降解以及切除嘧啶二聚体能力的检测。结果发现,核酸外切酶VI(polAex-)和核酸外切酶VII(xseA-)双缺陷的菌株在高温(43℃)下孵育时的存活能力显著降低,超过了之前观察到的polAex单突变体的降低程度。核酸外切酶VI缺陷菌株对UV和X射线的敏感性并没有因添加xseA7突变而增加。两种酶都缺陷的突变体在切除高达40 J/m2 UV产生的二聚体方面与野生型菌株效率相当。在更高剂量下,仅含有polAex-突变的菌株切除二聚体的能力降低;然而,在这些剂量下二聚体切除数据的解释因这些菌株照射后极端的DNA降解而变得复杂。polAex xseA7双突变体菌株额外的缺陷对照射后DNA降解或在polAex单突变体中观察到的高UV剂量下二聚体切除的明显缺陷均无显著影响。

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