Enhancement of frame-shift mutation by the overproduction of msDNA in Escherichia coli.
A minor population of wild Escherichia coli strains contain retroelements called retrons, which produce a peculiar satellite DNA, multicopy single-stranded DNA (msDNA). It has been reported that mismatched base pairs in the secondary structure formed in msDNA are mutagenic in E. coli[Maas et al.(1994) Mol.Microbiol. 14,437-441; Maas et al. (1996) Mol. Microbiol, 19, 505-509]. We reexamined this proposal by converting mismatched base pairs to matched base pairs using a single msDNA species, msDNA-Ec86, or by deleting mismatched regions using msDNA-Ec73. We also examined the effect of reverse transcriptases (RT) without msDNA production on mutagenesis. All the constructs are under the lpp/lac promoter-operator control so that their mutagenic effects can be tested in the absence and the presence of a lac inducer. It was found that when the production of msDNA-Ec86 or Ec73 was induced, reversion frequencies from Lac- to Lac+ significantly increased in the case of a Lac- mutation caused by a frame-shift mutation, but much less by a substitution mutation. The removal of mismatched base pairs eliminated the high mutation frequencies, and the inducible expression of RT alone was not mutagenic. These results are consistent with the hypothesis of Maas and his associates that mismatched base pairs in msDNA sequester a cellular mismatch repair system, resulting in the increase of frame-shift mutations.
Mao
J R
JR
Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Inouye
S
S
Inouye
M
M
D001482:Base Composition; D001483:Base Sequence; D004260:DNA Repair; D004261:DNA Replication; D004269:DNA, Bacterial / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D016368:Frameshift Mutation; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009153:Mutagens; D009690:Nucleic Acid Conformation; D018626:Retroelements / Q000235:genetics
D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D009153:Mutagens; D018626:Retroelements