Poly amine Requirement for Streptomycin Action on Protein Synthesis in Bacteria

Goldemberg, Sara H.; Algranati, Israel D.

doi:10.1111/j.1432-1033.1981.tb06330.x

Cited by 34 publications

(29 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…They are involved in macromolecular syntheses and in particular in the modulation of translation accuracy at steps which may be essential for survival of the cell populations (5). Polyamine auxotrophy in E. coli due to SpeD inactivation has been implicated in resistance to aminoglycoside antibiotics, including kasugamycin (9,10). Ribosomes of a polyamine auxotrophic E. coli mutant starved of polyamines showed a reduced affinity for streptomycin, and their protein synthesis activity was less sensitive to the drug (10).…”

Section: Discussionmentioning

confidence: 99%

Inactivation of KsgA, a 16S rRNA Methyltransferase, Causes Vigorous Emergence of Mutants with High-Level Kasugamycin Resistance

Ochi

Kim

Tanaka

et al. 2009

Antimicrob Agents Chemother

View full text Add to dashboard Cite

The methyltransferases RsmG and KsgA methylate the nucleotides G535 (RsmG) and A1518 and A1519 (KsgA) in 16S rRNA, and inactivation of the proteins by introducing mutations results in acquisition of low-level resistance to streptomycin and kasugamycin, respectively. In a Bacillus subtilis strain harboring a single rrn operon (rrnO), we found that spontaneous ksgA mutations conferring a modest level of resistance to kasugamycin occur at a high frequency of 10 ؊6 . More importantly, we also found that once cells acquire the ksgA mutations, they produce high-level kasugamycin resistance at an extraordinarily high frequency (100-fold greater frequency than that observed in the ksgA ؉ strain), a phenomenon previously reported for rsmG mutants. This was not the case for other antibiotic resistance mutations (Tsp r and Rif r ), indicating that the high frequency of emergence of a mutation for high-level kasugamycin resistance in the genetic background of ksgA is not due simply to increased persistence of the ksgA strain. Comparative genome sequencing showed that a mutation in the speD gene encoding S-adenosylmethionine decarboxylase is responsible for the observed high-level kasugamycin resistance. ksgA speD double mutants showed a markedly reduced level of intracellular spermidine, underlying the mechanism of high-level resistance. A growth competition assay indicated that, unlike rsmG mutation, the ksgA mutation is disadvantageous for overall growth fitness. This study clarified the similarities and differences between ksgA mutation and rsmG mutation, both of which share a common characteristic-failure to methylate the bases of 16S rRNA. Coexistence of the ksgA mutation and the rsmG mutation allowed cell viability. We propose that the ksgA mutation, together with the rsmG mutation, may provide a novel clue to uncover a still-unknown mechanism of mutation and ribosomal function.

show abstract

Section: Discussionmentioning

confidence: 99%

Inactivation of KsgA, a 16S rRNA Methyltransferase, Causes Vigorous Emergence of Mutants with High-Level Kasugamycin Resistance

Ochi

Kim

Tanaka

et al. 2009

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…Adenovirus proteins made in vitro in the presence of polyamines resembled more closely those synthesized in vivo, both in size and quantity [9]. Polypeptides of higher molecular mass were found in smaller amounts in the absence of polyamines in vitro [lo, 111 and in vivo [12]. These observations had suggested that polyamines might prevent premature peptide termination or otherwise affect the fidelity of translation.…”

mentioning

confidence: 96%

Section: Bacterial Strains and Bacteriophagementioning

confidence: 99%

“…Genetic manipulations were performed as described [21,221. Our polyamine-requiring strain BGA8 [spell spec thr leu thi luc sup (unknown amber suppressor)] has been previously described [12]. A his mutant of BGA8 was isolated following ethyl methanesulfonate mutagenesis ; the strain was made sensitive to MS2 by conjugation with BW104 (which contains F42 = F'lac') and selection on lactose.…”

Section: Bacterial Strains and Bacteriophagementioning

confidence: 99%

See 1 more Smart Citation

Effect of polyamines on translation fidelity in vivo

McMurry

Algranati

1986

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

Polyamines, when added to cell-free protein-synthesizing systems, have been shown either to reduce mistranslation or to increase it depending upon the composition of the reaction mixture. To study this question under conditions as natural as possible we investigated the effects of polyamines on the fidelity of protein synthesis in intact Escherichiu coli bacterial cells, using strains which were auxotrophic for polyamine synthesis. Error was measured in two ways: (a) the incorporation of [3H]histidine into coat protein of bacteriophage MS2, the gene of which does not code for histidine, and (b) the synthesis of a basic variant of MS2 coat protein in which a lysine erroneously replaces an asparagine, causing a change in isoelectric point. We found that when cell cultures were supplemented with polyamines there was no effect on the first type of error and the second type decreased twofold. The measured errors occurred at the level of translation because their frequency increased in the presence of streptomycin and decreased in cells bearing a streptomycin-resistance mutation known to lower the occurrence of translational misreading. The average erroneous incorporation per mol coat protein in the presence of polyamines was 1.43 ? 0.59 mmol histidine and 25 -34 mmol lysine/asparagine substitution. The reason for the different effect of polyamines on the two types of error is not known but could be related to the difference between their corresponding frequencies or to codon-specific effects.Polyamines are widely distributed organic cations which appear to have many roles, especially in processes involving nucleic acids, such as DNA replication, transcription and translation [I -51. Studies carried out with cell-free systems have demonstrated that polyamines can replace part of the magnesium requirement of protein synthesis and at the same time cause overall stimulation [6 -81. Polyamines have also been shown to have a qualitative effect on the proteins synthesized. Adenovirus proteins made in vitro in the presence of polyamines resembled more closely those synthesized in vivo, both in size and quantity [9]. Polypeptides of higher molecular mass were found in smaller amounts in the absence of polyamines in vitro [lo, 111 and in vivo [12]. These observations had suggested that polyamines might prevent premature peptide termination or otherwise affect the fidelity of translation. It was subsequently reported that polyamines, particularly spermidine, decreased the misincorporation of leucine for phenylalanine severalfold in a polyuridylic-acid-dependent in vitro translation system [13-151. However, in these studies the error determinations in presence of polyamines were done at lower Mg concentrations than those in the absence of polyamines because the Mg optimum for polyphenylalanine synthesis was higher under the latter conditions than in the presence of polyamines. In one study, other changes besides polyamine addition were made [16]. At some concentrations of Mg, however, the error was no different or [18] that when...

show abstract

“…In work with streptomycin-sensitive polyamineauxotrophic mutants we have shown that streptomycin caused a marked inhibition of protein synthesis in polyamine-supplemented cells, whereas bacteria starved for polyamines were less sensitive to the inhibitory action of the antibiotic [14]. These results might be due to the presence of defective 30 S subunits in bacteria submitted to polyamine starvation [ 15,16].…”

mentioning

confidence: 63%