Doyle N, Strike P
University of Liverpool, Department of Genetics and Microbiology, Donnan Laboratories, UK.
Mol Gen Genet. 1995 Jun 25;247(6):735-41. doi: 10.1007/BF00290405.
We have used the lacZ reversion assay to study the mutation spectra induced by the Escherichia coli chromosomal umuDC operon and of its two plasmid-borne analogues impCAB and mucAB following exposure of cells to UV light and methyl methanesulfonate (MMS). We have shown that the impCAB, mucAB and umuDC operons all produce a similar response to UV light which results almost exclusively in AT-->GC transitions. However, we found that the three operons produced different responses to alkylating agents. We found that with MMS the chromosomal umuDC operon produced almost exclusively AT-->GC transitions, whilst both mucAB and impCAB produced predominantly transversions. In the case of the impCAB operon the mutation spectrum contained more AT-->TA than GC-->TA transversions; this balance was reversed with mucAB. The effect of the copy number of the error-prone DNA repair operons upon the mutagenic spectra was also studied. The results obtained suggest that the copy number of the imp operon does not greatly affect the specificity of base substitutions observed. However, an increase in the copy number of the umuDC operon greatly affected the specificity of base substitution, such that virtually no transitions were produced and the spectrum was dominated by GC/AT-->TA transversions. It appears that the three error-prone DNA repair operons impCAB, mucAB and umuDC, despite showing strong structural and functional homologies, can display major differences in the spectrum of base changes induced during mutagenesis.(ABSTRACT TRUNCATED AT 250 WORDS)
我们已使用lacZ回复突变试验来研究大肠杆菌染色体umuDC操纵子及其两个质粒携带类似物impCAB和mucAB在细胞暴露于紫外线和甲基磺酸甲酯(MMS)后诱导产生的突变谱。我们已表明,impCAB、mucAB和umuDC操纵子对紫外线均产生类似反应,几乎完全导致AT→GC转换。然而,我们发现这三个操纵子对烷化剂产生不同反应。我们发现,对于MMS,染色体umuDC操纵子几乎完全导致AT→GC转换,而mucAB和impCAB均主要产生颠换。就impCAB操纵子而言,突变谱中AT→TA转换比GC→TA转换更多;而mucAB的情况则相反。还研究了易错DNA修复操纵子的拷贝数对诱变谱的影响。所得结果表明,imp操纵子的拷贝数对观察到的碱基替换特异性影响不大。然而,umuDC操纵子拷贝数的增加极大地影响了碱基替换的特异性,几乎不产生转换,且谱主要由GC/AT→TA颠换主导。看来,尽管易错DNA修复操纵子impCAB、mucAB和umuDC显示出强烈的结构和功能同源性,但在诱变过程中诱导产生的碱基变化谱中仍可表现出重大差异。(摘要截短于250字)