Slechta E Susan, Liu Jing, Andersson Dan I, Roth John R
Department of Biology, University of Utah, Salt Lake City, UT 84112, USA.
Genetics. 2002 Jul;161(3):945-56. doi: 10.1093/genetics/161.3.945.
In the genetic system of Cairns and Foster, a nongrowing population of an E. coli lac frameshift mutant appears to specifically accumulate Lac(+) revertants when starved on medium including lactose (adaptive mutation). This behavior has been attributed to stress-induced general mutagenesis in a subpopulation of starved cells (the hypermutable state model). We have suggested that, on the contrary, stress has no direct effect on mutability but favors only growth of cells that amplify their leaky mutant lac region (the amplification mutagenesis model). Selection enhances reversion primarily by increasing the mutant lac copy number within each developing clone on the selection plate. The observed general mutagenesis is attributed to a side effect of growth with an amplification-induction of SOS by DNA fragments released from a tandem array of lac copies. Here we show that the S. enterica version of the Cairns system shows SOS-dependent general mutagenesis and behaves in every way like the original E. coli system. In both systems, lac revertants are mutagenized during selection. Eliminating the 35-fold increase in mutation rate reduces revertant number only 2- to 4-fold. This discrepancy is due to continued growth of amplification cells until some clones manage to revert without mutagenesis solely by increasing their lac copy number. Reversion in the absence of mutagenesis is still dependent on RecA function, as expected if it depends on lac amplification (a recombination-dependent process). These observations support the amplification mutagenesis model.
在凯恩斯和福斯特的遗传系统中,当在含有乳糖的培养基上饥饿培养时,大肠杆菌乳糖移码突变体的非生长群体似乎会特异性地积累乳糖(+)回复突变体(适应性突变)。这种行为被归因于饥饿细胞亚群中应激诱导的普遍诱变(超可变状态模型)。相反,我们认为应激对突变率没有直接影响,而仅有利于扩增其渗漏突变型乳糖区域的细胞生长(扩增诱变模型)。选择主要通过增加选择平板上每个发育克隆内的突变型乳糖拷贝数来增强回复突变。观察到的普遍诱变归因于生长的副作用,即由乳糖拷贝串联阵列释放的DNA片段对SOS的扩增诱导。在这里,我们表明凯恩斯系统的肠炎沙门氏菌版本显示出依赖SOS的普遍诱变,并且在各方面的表现都与原始的大肠杆菌系统相似。在这两个系统中,乳糖回复突变体在选择过程中发生诱变。将突变率提高35倍的情况消除后,回复突变体数量仅减少2至4倍。这种差异是由于扩增细胞持续生长,直到一些克隆仅通过增加其乳糖拷贝数就能在不发生诱变的情况下回复突变。如预期的那样,如果回复突变依赖于乳糖扩增(一个依赖重组的过程),那么在没有诱变的情况下的回复突变仍然依赖于RecA功能。这些观察结果支持扩增诱变模型。