Influence of ciprofloxacin and vancomycin on mutation rate and transposition of IS256 in Staphylococcus aureus.
In Staphylococcus aureus, the development of intermediate resistance to vancomycin is due to an accumulation of mutations. To elucidate the mechanisms involved here, a standard laboratory strain (S. aureus HG001) and a clinical MRSA mutator strain (S. aureus SA1450/94, which is characterized by a spontaneous insertion of IS256 into the gene of the mismatch repair enzyme MutS) were incubated at subinhibitory concentrations of ciprofloxacin and vancomycin. Ciprofloxacin increased the mutation rates of both strains, but this effect was inhibited when the SOS response was blocked by the presence of a non-cleavable variant of the LexA repressor. In the presence of vancomycin, the mutation rate was slightly elevated in the mutator strain, and this increase also depended on the strain's ability to induce the SOS response. Furthermore, treatment with subinhibitory concentrations of both antibiotics resulted in an activation of transposition frequency of the insertion element IS256 in S. aureus HG001. Transposition was dependent on the presence of a functional transposase, and the activation of transposition depended on the presence of the functional phosphatase RsbU, which activates SigB transcription activity. An in silico analysis indicated a putative antisense sigma B promoter sequence within the transposase gene. Scrambling of this promoter resulted in an about 20-fold activation of transposition activity of IS256. These data indicate that sigma B is involved in the regulation of IS256 activity by generation of an antisense RNA.
Institute of Medical Microbiology, Immunology and Parasitology, University of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany.
Michael
M
Nagel
Tina
T
Reuter
Andrea
A
Jansen
Christiane
C
Szekat
Gabriele
G
Bierbaum
D000900:Anti-Bacterial Agents / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D002939:Ciprofloxacin / Q000378:metabolism*; D004251:DNA Transposable Elements / Q000187:drug effects*; D009154:Mutation; D011995:Recombination, Genetic; D013014:SOS Response, Genetics; D012808:Sigma Factor / Q000378:metabolism; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000235:genetics; D014640:Vancomycin / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C079011:SigB protein, Bacteria; D012808:Sigma Factor; D002939:Ciprofloxacin; D014640:Vancomycin