Effect of Serine Hydroxamate on the Growth of
            <i>Escherichia coli</i>

Tosa, Tetsuya; Pizer, Lewis I.

doi:10.1128/jb.106.3.966-971.1971

Cited by 56 publications

(43 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The model predicts, therefore, that the same degree of inhibition of serine‐tRNA synthetase should stimulate bypassing much more in the UCC construct than in the AGC construct. The charging of all serine‐tRNA isoacceptors was inhibited with the analogue serine‐hydroxamate (SHX; Tosa & Pizer, 1971; Pizer & Merlie, 1973). The prediction was confirmed by a qualitative, visual test of the analogue's effect on β‐galactosidase synthesis encoded by the two reporters (Fig 1).…”

Section: Resultsmentioning

confidence: 99%

Ribosome bypassing at serine codons as a test of the model of selective transfer RNA charging

et al. 2005

View full text Add to dashboard Cite

Recently, a model of the flux of amino acids through transfer RNAs (tRNAs) and into protein has been developed. The model predicts that the charging level of different isoacceptors carrying the same amino acid respond very differently to variation in supply of the amino acid or of the rate of charging. It has also been shown that ribosome bypassing is specifically stimulated at 'hungry' codons calling for an aminoacyl-tRNA in short supply. We have constructed two reporters of bypassing, which differ only in the identity of the serine codon subjected to starvation. The stimulation of bypassing as a function of starvation differed greatly between the two serine codons, in good agreement with the quantitative predictions of the model. Keywords: amino acid; bypassing; charging; protein synthesis; tRNA EMBO reports (2005) 6, 147-150. doi:10.1038/sj.embor.7400332 Elf et al (2003) have developed a theoretical model of the response to amino-acid limitation of the different members (isoacceptors) of each family of transfer RNAs (tRNAs) that carry the same amino acid. The theory rests on considerations of supply and demand. The parameters of supply are the rate of synthesis of the limiting amino acid and the abundance of each isoacceptor tRNA in question; the parameters of demand are the frequencies with which the codons read by a given isoacceptor are found in translating ribosomes, as well as the overall capacity of ribosomes to consume the amino acid. The model leads to the counterintuitive prediction that amino-acid limitation can elicit very different degrees of residual aminoacyl charging among the different isoacceptors. Those that have a high concentration relative to the frequency of their cognate codons in messenger RNAs will retain a much higher level of charging than those in which the ratio is lower. We report here a test of this prediction in the case of two serine-tRNA isoacceptors, using ribosome bypassing as a reflection of the starvation experienced by codons calling for the two different isoacceptors. INTRODUCTION RESULTS AND DISCUSSIONIt has been demonstrated elsewhere that 'hungry' codons calling for a tRNA, the acylated form of which is in short supply, are the sites of reading frame errors Peter et al, 1992;Lindsley & Gallant, 1993;Barak et al, 1996;Gallant et al, 2000) and ribosome bypassing (Gallant & Lindsley, 1998;Gallant et al, 2003;Lindsley et al, 2003). Furthermore, minigene experiments have indicated that it is the absence of the aminoacylated tRNA, rather than any positive effect of the deacylated species, that gives rise to these ribosome errors . Therefore, the stimulation of either one of these ribosome gymnastic feats at a hungry codon should reflect the reduced charging level of that codon's cognate tRNA, an analytic approach that has been introduced already in the case of isoleucine tRNAs (Kaplun et al, 2002;Elf et al, 2003).To test the model of selective tRNA charging, we have constructed reporters of bypassing, which differ only at the serine codon subjected to starvation. The repo...

show abstract

Section: Resultsmentioning

confidence: 99%

Ribosome bypassing at serine codons as a test of the model of selective transfer RNA charging

et al. 2005

View full text Add to dashboard Cite

show abstract

“…The ancestor and 17 000‐generation S and L strains were streaked onto LB plates. After 1 day, cells were harvested from the plates and re‐suspended in MOPS medium containing glucose as the sole carbon source and serine hydroxamate (0.2 mg mL −1 ) to artificially induce the stringent response (Tosa & Pizer 1971). At time zero, samples were diluted 25‐fold in the same medium containing [32P] H 3 PO 4 (10 μCi mL −1 ).…”

Section: Methodsmentioning

confidence: 99%

Death and cannibalism in a seasonal environment facilitate bacterial coexistence

Rozen¹,

Philippe²,

Visser³

et al. 2008

Ecology Letters

119

136

View full text Add to dashboard Cite

Bacterial populations can evolve and adapt to become diverse niche specialists, even in seemingly homogeneous environments. One source of this diversity arises from newly 'constructed' niches that result from the activities of the bacteria themselves. Ecotypes specialized to exploit these distinct niches can subsequently coexist via frequency-dependent interactions. Here, we describe a novel form of niche construction that is based upon differential death and cannibalism, and which evolved during 20 000 generations of experimental evolution in Escherichia coli in a seasonal environment with alternating growth and starvation. In one of 12 populations, two monophyletic ecotypes, S and L, evolved that stably coexist with one another. When grown and then starved in monoculture, the death rate of S exceeds that of L, whereas the reverse is observed in mixed cultures. As shown by experiments and numerical simulations, the competitive advantage of S cells is increased by extending the period of starvation, and this advantage results from their cannibalization of the debris of lysed L cells, which allows the S cells to increase both their growth rate and total cell density. At the molecular level, the polymorphism is associated with divergence in the activity of the alternative sigma factor RpoS, with S cells displaying no detectable activity, while L cells show increased activity relative to the ancestral genotype. Our results extend the repertoire of known cross-feeding mechanisms in microbes to include cannibalism during starvation, and confirm the central roles for niche construction and seasonality in the maintenance of microbial polymorphisms.

show abstract

“…SHX is permeable and inhibits binding of Serine to the seryl-tRNA synthetase. This treatment is known to induce synthesis of (p)ppGpp in E. coli (Tosa and Pizer, 1971).…”

Section: Time Dependence Of Cumulative (P)ppgpp Production In Strainsmentioning

confidence: 99%

“…SHX is permeable and prevents charging of tRNA Ser by seryl-tRNA synthetase (Tosa and Pizer, 1971). This treatment is known to induce synthesis of (p)ppGpp in E. coli.…”

Section: Tests Of Labeled Rela: Intracellular Nucleotide Quantificatimentioning

confidence: 99%

Effects of amino acid starvation on RelA diffusive behavior in live Escherichia coli

Bouveret

Zhang

et al. 2015

Molecular Microbiology

View full text Add to dashboard Cite

SummaryDuring amino acid starvation, bacterial cells rapidly synthesize the nucleotides (p)ppGpp, causing a massive re-programming of the transcriptional profile known as the stringent response. The (p)ppGpp synthase RelA is activated by ribosomes harboring an uncharged tRNA at the A site. It is unclear whether synthesis occurs while RelA is bound to the ribosome or free in the cytoplasm. We present a study of three Escherichia coli strains, each expressing a different RelA-fluorescent protein (RelA-FP) construct: RelA-YFP, RelA-mEos2 and RelA-Dendra2. Single-molecule localization and tracking studies were carried out under normal growth conditions and during amino acid starvation. Study of three labeling schemes enabled us to assess potential problems with FP labeling of RelA. The diffusive trajectories and axial spatial distributions indicate that amino acid starvation induces net binding of all three RelA-FP constructs to 70S ribosomes. The data are most consistent with a model in which RelA synthesizes (p)ppGpp while bound to the 70S ribosome. We suggest a 'short hopping time' model of RelA activity during starvation. Our results contradict an earlier study of RelA-Dendra2 diffusion that inferred offribosome synthesis of (p)ppGpp. The reasons for the discrepancy remain unclear.

show abstract

Effect of Serine Hydroxamate on the Growth of Escherichia coli

Cited by 56 publications

References 15 publications

Ribosome bypassing at serine codons as a test of the model of selective transfer RNA charging

Ribosome bypassing at serine codons as a test of the model of selective transfer RNA charging

Death and cannibalism in a seasonal environment facilitate bacterial coexistence

Effects of amino acid starvation on RelA diffusive behavior in live Escherichia coli

Contact Info

Product

Resources

About