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枯草芽孢杆菌中DNA修复与转化的进化。II. 诱导修复的作用。

DNA repair and the evolution of transformation in Bacillus subtilis. II. Role of inducible repair.

作者信息

Wojciechowski M F, Hoelzer M A, Michod R E

机构信息

Department of Ecology and Evolutionary Biology, University of Arizona, Tucson 85721.

出版信息

Genetics. 1989 Mar;121(3):411-22. doi: 10.1093/genetics/121.3.411.

Abstract

In Bacillus subtilis, DNA repair and recombination are intimately associated with competence, the physiological state in which the bacterium can bind, take up and recombine exogenous DNA. Previously, we have shown that the homologous DNA transformation rate (ratio of transformants to total cells) increases with increasing UV dosage if cells are transformed after exposure to UV radiation (UV-DNA), whereas the transformation rate decreases if cells are transformed before exposure to UV (DNA-UV). In this report, by using different DNA repair-deficient mutants, we show that the greater increase in transformation rate in UV-DNA experiments than in DNA-UV experiments does not depend upon excision repair or inducible SOS-like repair, although certain quantitative aspects of the response do depend upon these repair systems. We also show that there is no increase in the transformation rate in a UV-DNA experiment when repair and recombination proficient cells are transformed with nonhomologous plasmid DNA, although the results in a DNA-UV experiment are essentially unchanged by using plasmid DNA. We have used din operon fusions as a sensitive means of assaying for the expression of genes under the control of the SOS-like regulon in both competent and noncompetent cell subpopulations as a consequence of competence development and our subsequent experimental treatments. Results indicate that the SOS-like system is induced in both competent and noncompetent subpopulations in our treatments and so should not be a major factor in the differential response in transformation rate observed in UV-DNA and DNA-UV treatments. These results provide further support to the hypothesis that the evolutionary function of competence is to bring DNA into the cell for use as template in the repair of DNA damage.

摘要

在枯草芽孢杆菌中,DNA修复和重组与感受态密切相关,感受态是细菌能够结合、摄取并重组外源DNA的生理状态。此前,我们已经表明,如果细胞在紫外线辐射(UV-DNA)后进行转化,同源DNA转化率(转化子与总细胞数的比率)会随着紫外线剂量的增加而增加;而如果细胞在紫外线照射前进行转化(DNA-UV),转化率则会降低。在本报告中,通过使用不同的DNA修复缺陷突变体,我们表明,UV-DNA实验中转化率的增加幅度大于DNA-UV实验,这并不依赖于切除修复或诱导型SOS样修复,尽管反应的某些定量方面确实依赖于这些修复系统。我们还表明,当用非同源质粒DNA转化修复和重组能力正常的细胞时,UV-DNA实验中的转化率不会增加,尽管使用质粒DNA时DNA-UV实验的结果基本不变。由于感受态的发展以及我们随后的实验处理,我们使用din操纵子融合作为一种灵敏的方法,来检测在感受态和非感受态细胞亚群中,受SOS样调控子控制的基因的表达。结果表明,在我们的处理中,SOS样系统在感受态和非感受态亚群中均被诱导,因此它不应是在UV-DNA和DNA-UV处理中观察到的转化率差异反应的主要因素。这些结果为以下假设提供了进一步的支持,即感受态的进化功能是将DNA带入细胞,用作修复DNA损伤的模板。

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