Watanabe-Akanuma M, Woodgate R, Ohta T
Institute of Environmental Toxicology, Tokyo, Japan.
Mutat Res. 1997 Jan 3;373(1):61-6. doi: 10.1016/s0027-5107(96)00189-3.
RecA730 belongs to a class of mutant RecA protein that is often referred to as RecA*, since it is constitutively activated for coprotease functions in the absence of exogenous DNA-damage. Escherichia coli strains carrying recA730 (or other recA* alleles) exhibit dramatic increases in SOS-dependent spontaneous mutator activity. We have analyzed the specificity of this mutator phenotype by employing F'-plasmids carrying a set of mutant lacZ genes that can individually detect two types of transitions, four types of transversions, and four kinds of specific frameshift events. Analysis revealed that most of the spontaneous mutagenesis in a recA730 lexA51(Def) strain (which expresses derepressed levels of all LexA-regulated proteins) can be attributed to a specific increase in A:T-->T:A, A:T-->C:G and G:C-->T:A transversions, with the A:T-->T:A transversions occurring most frequently. These transversion events were completely abolished in a delta umuDC strain, indicating that the functionally active UmuD'C proteins are normally required for their generation. The spectrum obtained was similar to that of strains with a defect in the epsilon (3'-->5' proofreading) subunit of DNA polymerase III. Such an observation raises the possibility that the wild-type epsilon protein is in activated in strains expressing the RecA730 and UmuD'C proteins.
RecA730属于一类突变型RecA蛋白,通常被称为RecA*,因为在没有外源DNA损伤的情况下,它的共蛋白酶功能被组成型激活。携带recA730(或其他recA*等位基因)的大肠杆菌菌株在SOS依赖的自发诱变活性上表现出显著增加。我们通过使用携带一组突变lacZ基因的F'-质粒分析了这种诱变表型的特异性,这些突变lacZ基因可以分别检测两种类型的转换、四种类型的颠换和四种特定的移码事件。分析表明,recA730 lexA51(Def)菌株(表达所有LexA调控蛋白的去阻遏水平)中的大多数自发诱变可归因于A:T-->T:A、A:T-->C:G和G:C-->T:A颠换的特异性增加,其中A:T-->T:A颠换发生得最频繁。这些颠换事件在ΔumuDC菌株中完全消失,表明功能活跃的UmuD'C蛋白通常是其产生所必需的。所获得的谱与DNA聚合酶III的ε(3'-->5'校对)亚基有缺陷的菌株相似。这样的观察结果增加了一种可能性,即野生型ε蛋白在表达RecA730和UmuD'C蛋白的菌株中被激活。