Yi T M, Stearns D, Demple B
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
J Bacteriol. 1988 Jul;170(7):2898-903. doi: 10.1128/jb.170.7.2898-2903.1988.
We studied DNA rearrangements in Escherichia coli by using a plasmid-based system with a transcriptionally silent tet gene and selecting for Tetr isolates. The predominant activating event was a 1.3-kilobase-pair deletion in the plasmid between two sites, with 14 of 19 base pairs being identical. These deletions occurred equally frequently in a recA+ strain and a recA13 mutant. However, the frequency of Tetr occurrence was stimulated 50-fold by treatment of the cells with UV light in a process that was at least partly independent of the SOS response. Bacterial mutants deleted for the xth-pnc region of the chromosome exhibited a strongly elevated spontaneous frequency of Tetr isolates, all with the same 1.3-kilobase-pair deletion. This phenotype of high-frequency deletion could be complemented by an episome covering this region, but not by the cloned xth gene. These studies helped to define the role of different DNA damages in illegitimate recombination and identify a region of the E. coli chromosome that contains a gene whose product normally suppresses illegitimate deletions.
我们通过使用一个基于质粒的系统来研究大肠杆菌中的DNA重排,该系统带有一个转录沉默的tet基因,并筛选Tetr分离株。主要的激活事件是质粒中两个位点之间发生了1.3千碱基对的缺失,其中19个碱基对中有14个是相同的。这些缺失在recA+菌株和recA13突变体中发生的频率相同。然而,用紫外线处理细胞会使Tetr出现的频率提高50倍,这一过程至少部分独立于SOS反应。缺失染色体xth-pnc区域的细菌突变体表现出Tetr分离株的自发频率大幅升高,所有突变体都有相同的1.3千碱基对缺失。高频缺失的这种表型可以由覆盖该区域的附加体互补,但不能由克隆的xth基因互补。这些研究有助于确定不同DNA损伤在非法重组中的作用,并鉴定出大肠杆菌染色体上一个包含其产物通常抑制非法缺失的基因的区域。