Autostimulation of Dihydrostreptomycin Uptake in Bacillus subtilis

SUMMARY: In Bacillus subtilis it was shown that the membrane potential (Δ,ψ) has to reach a threshold value of −180 to −190 mV for efficient uptake of dihydrostreptomycin to occur. The magnitude of Δ,ψ is raised above this threshold, and dihydrostreptomycin uptake greatly enhanced, not only by dihydrostreptomycin itself (autostimulation) and by other miscoding aminoglycoside antibiotics, but also by puromycin, bacitracin and N, N-dicyclohexylcarbodiimide. Stimulation of uptake by dihydrostreptomycin or puromycin was dependent on a specific interference with ongoing protein synthesis. Thus, chloramphenicol prevented the stimulating effect of puromycin by lowering the magnitude of δ+. Although normally severely antagonizing streptomycin accumulation, K+, as well as spermidine and putrescine, which are known to stabilize ribosomes, consequently enhanced autostimulation of dihydrostreptomycin uptake in a K+-retention mutant with impaired protein synthesis. It is suggested that miscoding aminoglycosides and puromycin both enhance dihydrostreptomycin uptake by increasing δ+ due to ion fluxes, which are themselves caused by a dramatic stimulation of intracellular proteolysis of faulty proteins.