Effect of polyamines on plasmid-mediated kanamycin resistance and kanamycin phosphotransferase gene expression in Escherichia coli.

The emergence of kanamycin resistance in a polyamine-deficient mutant of E. coli transformed with a plasmid encoding the kanamycin phosphotransferase gene has been studied. The initial inhibition of growth and protein synthesis caused by the addition of the antibiotic could be reversed earlier in polyamine-supplemented bacteria than in those depleted of the organic bases. Concomitantly, we have observed that the increase of kanamycin phosphotransferase activity evoking the antibiotic resistance was higher in bacteria cultivated in the presence of putrescine. This result seems to depend exclusively on the enhanced capacity of the translation process in bacteria grown with polyamines since the transcription of phosphotransferase gene was higher in cells subjected to polyamine starvation.

Polyamines modulate streptomycin-induced mistranslation in Escherichia coli.

The effects of intracellular levels of polyamines on both the in vivo inhibition of protein synthesis and the decrease of translation accuracy induced by streptomycin have been studied in polyamine-auxotrophic strains of Escherichia coli infected with the MS2 bacteriophage. The amount of viral coat protein formed was strongly reduced upon addition of increasing concentrations of streptomycin to polyamine-supplemented bacteria. In contrast, the antibiotic almost did not inhibit coat protein synthesis in polyamine-starved cells. The increase of mistranslation frequency elicited by streptomycin was only observed in bacteria grown with putrescine. In these cells several coat protein-satellites were detected after two-dimensional gel electrophoresis. These proteins, more basic than the normal MS2 coat protein, contain multiple substitutions of lysine for asparagine.

Inhibition of protein synthesis by aminoglycoside antibiotics in polyamine-requiring bacteria.

The effect of streptomycin and other aminoglycosides on protein synthesis has been studied using various streptomycin-sensitive strains unable to synthesize polyamines. We have confirmed and extended our previous results showing that the strong inhibition of translation caused by the antibiotic in polyamine-supplemented bacteria was markedly reduced in polyamine-starved cells. The analysis of polypeptides synthesized in the absence and presence of streptomycin in bacteria grown with and without putrescine has shown that the antibiotic provoked the accumulation of low molecular weight peptides partially bound to ribosomes in polyamine-unstarved cells. On the contrary, the drug did not induce major alterations in the patterns of proteins obtained from polyamine-depleted bacteria. The addition of the antibiotic did not evoke any change of proteolytic activity.

Protein synthesis in polyamine-deficient bacteria during amino-acid starvation.

Amino-acid starvation in polyamine-auxotrophic bacteria grown in the presence of putrescine provokes a marked inhibition of protein synthesis. This inhibition is almost completely relieved in polyamine-depleted cells. The differential behaviour of bacterial protein synthesis depending on the endogenous levels of polyamines is not due to a change in the uptake of amino acids used to measure protein synthesis, nor to the decreased growth rate of polyamine-depleted cells. During leucine starvation, cells grown with putrescine synthesized a somewhat lower amount of high-molecular-weight proteins than polyamine-depleted bacteria. In addition, cells with normal endogenous levels of polyamines accumulated significant amounts of 62 and 41 kDa polypeptides as well as several low-molecular-weight peptides.