Maas W K, Wang C, Lima T, Hach A, Lim D
Department of Microbiology, New York University Medical Center, New York USA.
Mol Microbiol. 1996 Feb;19(3):505-9. doi: 10.1046/j.1365-2958.1996.392921.x.
Multicopy single-stranded DNA (msDNA) molecules consist of single-stranded DNA covalently linked to RNA. In Escherichia coli, such molecules are encoded by genetic elements called retrons. The DNA moieties of msDNAs have characteristic stem-loop structures, and most of these structures contain mismatched base pairs. Previously, we showed that retrons encoding msDNAs with mismatched base pairs are mutagenic when present in multicopy plasmids. In this study we show that such msDNAs, in a similar manner to genetic defects in mismatch repair, increase the frequency of interspecies recombination in matings between Salmonella typhimurium and E. coli. To demonstrate interference with mismatch repair by msDNA, we show that the addition of a plasmid containing the gene for MutS protein suppresses the mutagenic and recombinogenic effects of msDNAs. We also show that in mutS mutants, msDNA does not increase the frequency of either mutations or interspecies recombination. We conclude from these findings that the mutagenic and recombinogenic effects of msDNAs are due to titrating out MutS protein.
多拷贝单链DNA(msDNA)分子由与RNA共价连接的单链DNA组成。在大肠杆菌中,此类分子由称为反转录子的遗传元件编码。msDNA的DNA部分具有特征性的茎环结构,并且这些结构中的大多数都含有错配碱基对。此前,我们发现,当存在于多拷贝质粒中时,编码带有错配碱基对的msDNA的反转录子具有致突变性。在本研究中,我们发现此类msDNA与错配修复中的遗传缺陷类似,会增加鼠伤寒沙门氏菌与大肠杆菌交配时种间重组的频率。为了证明msDNA对错配修复的干扰,我们发现添加含有MutS蛋白基因的质粒可抑制msDNA的致突变和重组效应。我们还发现,在mutS突变体中,msDNA不会增加突变或种间重组的频率。从这些发现中我们得出结论,msDNA的致突变和重组效应是由于消耗了MutS蛋白。