Error-prone DNA repair system in enteroaggregative Escherichia coli identified by subtractive hybridization.
Enteroaggregative Escherichia coli (EAEC) are etiologic agents of diarrhea. The EAEC category is heterogeneous, but most in-depth experimentation has focused on prototypical strain, 042. We hypothesized that 60A, another EAEC strain, might posses virulence or fitness genes that 042 does not have. Through subtractive hybridization we identified 60A-specific sequences, including loci present in other E. coli and phage DNA. One locus thus identified was impB, a LexA repressed error-prone DNA repair gene that has been identified in plasmids from other enteric organisms and which we detected in 21 of 34 EAEC strains. An isogenic 60A impB mutant showed decreased survival and mutagenesis after exposure to UV, as well as bile salt exposure, compared to the wild-type strain, and these phenotypes could be complemented in trans. The EAEC strain 60A imp operon differs structurally from previously described homologs. A cryptic gene, impC, present in other imp operons, is absent from 60A. In addition, transcription of impAB in strain 60A occurs from a promoter that is dissimilar to the previously described impC promoter but is still triggered by UV-mediated damage. In strain 60A the impAB and the aggregative adherence fimbriae I (AAF/I)-encoding genes are on the same large plasmid, and the 60A version of the operon is predominantly seen in AAF/I-positive EAEC. Supplementary imp SOS-inducible error-prone repair systems are common among EAEC even though they are absent in prototypical strain 042.
Joo
Lucy M
LM
Department of Biology, Haverford College, 370 Lancaster Ave., Haverford, PA 19041, USA.
Macfarlane-Smith
Louissa R
LR
Okeke
Iruka N
IN
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000494:pharmacology; D001422:Bacterial Adhesion; D001647:Bile Acids and Salts / Q000494:pharmacology; D018441:Biofilms; D002939:Ciprofloxacin / Q000494:pharmacology; D004260:DNA Repair / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000472:pathogenicity / Q000528:radiation effects*; D029968:Escherichia coli Proteins / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods; D009876:Operon / Q000235:genetics; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D013307:Streptomycin / Q000494:pharmacology; D014466:Ultraviolet Rays; D014774:Virulence
D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D001647:Bile Acids and Salts; D029968:Escherichia coli Proteins; C489316:ImpB protein, E coli; D002939:Ciprofloxacin; D013307:Streptomycin