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ssb - 113等位基因抑制dnaQ49突变体,并改变大肠杆菌中的DNA超螺旋。

The ssb-113 allele suppresses the dnaQ49 mutator and alters DNA supercoiling in Escherichia coli.

作者信息

Quiñones A, Neumann S

机构信息

Institut für Genetik, Martin-Luther-Universität, Halle, Germany.

出版信息

Mol Microbiol. 1997 Jul;25(2):237-46. doi: 10.1046/j.1365-2958.1997.4531718.x.

Abstract

Mutations in the dnaQ gene, which encodes the proofreading epsilon-subunit of the DNA polymerase III holoenzyme, lead to a mutator phenotype caused by enhanced error rates during DNA replication. In this paper, we studied the influence of ssb mutations on the dnaQ49 mutator, because of the involvement of SSB protein in DNA replication. We found that the ssb-113 mutation suppresses the mutator phenotype of dnaQ49. The suppression effect resulted from an enhanced expression of the dnaQ49 allele as determined by experiments with gene fusions. S1 nuclease analysis revealed that the increased dnaQ expression is based on transcriptional activation of the dnaQP2 promoter. This seems to be the consequence of an increased DNA supercoiling in the ssb-113 mutant, which also influenced further functions that are sensitive to alterations in DNA supercoiling. These results support the hypothesis that the expression of the epsilon-subunit of DNA polymerase III may additionally be modulated by DNA supercoiling, and suggest a possible role for DNA topology in mutagenesis.

摘要

dnaQ基因发生突变会导致一种突变体表型,该基因编码DNA聚合酶III全酶的校对ε亚基,突变会使DNA复制过程中的错误率增加。在本文中,由于SSB蛋白参与DNA复制,我们研究了ssb突变对dnaQ49突变体的影响。我们发现,ssb - 113突变抑制了dnaQ49的突变体表型。通过基因融合实验确定,这种抑制作用源于dnaQ49等位基因表达的增强。S1核酸酶分析表明,dnaQ表达的增加是基于dnaQP2启动子的转录激活。这似乎是ssb - 113突变体中DNA超螺旋增加的结果,DNA超螺旋增加也影响了对DNA超螺旋改变敏感的其他功能。这些结果支持了以下假说:DNA聚合酶III的ε亚基的表达可能还受DNA超螺旋的调节,并提示了DNA拓扑结构在诱变中的可能作用。

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