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大肠杆菌DNA聚合酶I突变形式的生化特性。I. polA12突变

Biochemical characterization of mutant forms of DNA polymerase I from Escherichia coli. I. The polA12 mutation.

作者信息

Uyemura D, Lehman I R

出版信息

J Biol Chem. 1976 Jul 10;251(13):4078-84.

PMID:6470
Abstract

DNA polymerase I has been purified to greater than 90% homogeneity from a strain of Escherichia coli K12 that bears the temperature-sensitive DNA polymerase I mutatation, polA12. The mutant enzyme has a reduced electrophoretic mobility and sedimentation rate. It is abnormally thermolabile and is rapidly inactivated at low salt concentrations. Its polymerase and 5' leads to 3' exonuclease activities are not grossly defective at 30 degrees, yet its capacity to promote the concerted 5' leads to 3' polymerization and the 5' leads to 3' exonucleolytic hydrolysis of nucleotides at a nick ("nick translation") is decreased 10-fold. These effects are probably the result of a significant alteration in the tertiary structure of the enzyme.

摘要

已从携带温度敏感型DNA聚合酶I突变体polA12的大肠杆菌K12菌株中纯化出了纯度高于90%的DNA聚合酶I。该突变酶的电泳迁移率和沉降速率降低。它对热异常敏感，在低盐浓度下会迅速失活。其聚合酶活性和5'→3'外切核酸酶活性在30℃时并无严重缺陷，但其促进协同5'→3'聚合以及在切口处进行5'→3'核苷酸外切水解（“切口平移”）的能力下降了10倍。这些效应可能是该酶三级结构发生显著改变的结果。

相似文献

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Biochemical characterization of mutant forms of DNA polymerase I from Escherichia coli. I. The polA12 mutation.大肠杆菌DNA聚合酶I突变形式的生化特性。I. polA12突变
J Biol Chem. 1976 Jul 10;251(13):4078-84.
2
Biochemical characterization of mutant forms of DNA polymerase I from Escherichia coli. II. The polAex1 mutation.大肠杆菌DNA聚合酶I突变形式的生化特性。II. polAex1突变
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A conditional lethal mutant of Escherichia coli K12 defective in the 5' leads to 3' exonuclease associated with DNA polymerase I.大肠杆菌K12的一种条件致死突变体，其在与DNA聚合酶I相关的5'至3'核酸外切酶方面存在缺陷。
Proc Natl Acad Sci U S A. 1974 May;71(5):2048-51. doi: 10.1073/pnas.71.5.2048.
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Two temperature-sensitive polA mutants: an approach to the role in vivo of DNA polymerase I.两个温度敏感型polA突变体：探究DNA聚合酶I在体内作用的一种方法。
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Persistence of deoxyribonucleic acid polymerase I and its 5'--3' exonuclease activity in PolA mutants of Escherichia coli K12.大肠杆菌K12的PolA突变体中脱氧核糖核酸聚合酶I及其5'→3'核酸外切酶活性的持续性
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Some properties of mutants of Escherichia coli K12 defective in DNA polymerase I and exonuclease V.大肠杆菌K12中DNA聚合酶I和核酸外切酶V缺陷型突变体的一些特性
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Genetic analysis of a temperature-resistant revertant of the conditional lethal Escherichia coli double mutant polA12 uvrE502.条件致死型大肠杆菌双突变体polA12 uvrE502的温度抗性回复突变体的遗传分析。
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An antimutator deoxyribonucleic acid polymerase. I. Purification and properties of the enzyme.一种抗突变脱氧核糖核酸聚合酶。I. 酶的纯化及性质
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J Biol Chem. 1975 Jan 25;250(2):470-8.

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