SOS mutator activity: unequal mutagenesis on leading and lagging strands.
A major pathway of mutagenesis in Escherichia coli is mediated by the inducible SOS response. Current models of SOS mutagenesis invoke the interaction of RecA and UmuD'(2)C proteins with a stalled DNA replication complex at sites of DNA lesions or poorly extendable terminal mismatches, resulting in an (error-prone) continuation of DNA synthesis. The precise mechanisms of SOS-mediated lesion bypass or mismatch extension are not known. Here, we have studied mutagenesis on the E. coli chromosome in recA730 strains. In recA730 strains, the SOS system is expressed constitutively, resulting in a spontaneous mutator effect (SOS mutator) because of reduced replication fidelity. We investigated whether during SOS mutator activity replication fidelity might be altered differentially in the leading and lagging strand of replication. Pairs of recA730 strains were constructed differing in the orientation of the lac operon relative to the origin of replication. The strains were also mismatch-repair defective (mutL) to facilitate scoring of replication errors. Within each pair, a given lac sequence is replicated by the leading-strand machinery in one orientation and by the lagging-strand machinery in the other orientation. Measurements of defined lac mutant frequencies in such pairs revealed large differences between the two orientations. Furthermore, in all cases, the frequency bias was the opposite of that seen in normal cells. We suggest that, for the lacZ target used in this study, SOS mutator activity operates with very different efficiency in the two strands. Specifically, the lagging strand of replication appears most susceptible to the SOS mutator effect.
Maliszewska-Tkaczyk
M
M
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5A, 02-106 Warsaw, Poland.
Jonczyk
P
P
Bialoskorska
M
M
Schaaper
R M
RM
Fijalkowska
I J
IJ
D000483:Alleles; D020137:Base Pair Mismatch; D004261:DNA Replication; D004269:DNA, Bacterial / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D016296:Mutagenesis; D011938:Rec A Recombinases / Q000235:genetics*; D013014:SOS Response, Genetics
D004269:DNA, Bacterial; D011938:Rec A Recombinases