Aminoglycoside-resistant mutation of Pseudomonas aeruginosa defective in cytochrome c552 and nitrate reductase.
A gentamicin-resistant mutant of Pseudomonas aeruginosa PAO503 was selected after ethyl methane sulfonate mutagenesis. The strain, P. aeruginosa PAO2401 had increased resistance to all aminoglycosides tested but exhibited no change for other antibiotics. The mutation designated aglA (aminoglycoside resistance) was 50% cotransducible with the 8-min ilvB,C marker on the P. aeruginosa chromosome. It showed a marked reduction in cytochrome c(552) and nitrate reductase (Nar) and a change in terminal oxidase activity. Cytochrome c(552) is a component of the P. aeruginosa Nar. No changes in succinate and reduced nicotinamide adenine dinucleotide dehydrogenases, ubiquinone content, Mg(2+)/Ca(2+) membrane adenosine triphosphatase, and energy coupling of electron transport to adenosine 5'-triphosphate synthesis were detected. Transport of gentamicin and dihydrostreptomycin was impaired in PAO2401, but transport of proline, arginine, glutamine, glucose or the polyamine spermidine was not reduced. Ribosomes of PAO2401, and PAO503 bound dihydrostreptomycin equally well, and cell extracts did not inactivate gentamicin or dihydrostreptomycin. Strain PAO2401 is resistant to gentamicin and dihydrostreptomycin because of impaired transport of these compounds. The transport studies indicate a selective coupling of dihydrostreptomycin and gentamicin transport with terminal electron transport. This conclusion was supported by results from another mutant (PAO417-T2) with increased Nar activity, enhanced dihydrostreptomycin and gentamicin transport and a reduction in resistance to these drugs. These results are discussed in relation to a refined model for aminoglycoside transport and briefly relative to plasmid-mediated aminoglycoside resistance.
D000596:Amino Acids / Q000378:metabolism; D000617:Aminoglycosides / Q000378:metabolism / Q000494:pharmacology; D001692:Biological Transport; D002463:Cell Membrane Permeability; D003574:Cytochrome c Group / Q000032:analysis; D004352:Drug Resistance, Microbial; D004734:Energy Metabolism; D005839:Gentamicins / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D009154:Mutation; D009565:Nitrate Reductases / Q000032:analysis; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism; D013307:Streptomycin / Q000378:metabolism
D000596:Amino Acids; D000617:Aminoglycosides; D003574:Cytochrome c Group; D005839:Gentamicins; D009565:Nitrate Reductases; D005947:Glucose; D013307:Streptomycin