Studies on the Control of Development. Accumulation of Guanosine Tetraphosphate and Pentaphosphate in Response to Inhibition of Protein Synthesis in Bacillus subtilis
- 1 August 1975
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 56 (2), 385-392
- https://doi.org/10.1111/j.1432-1033.1975.tb02244.x
Abstract
Bacillus subtilis cells accumulate unusual phosphorylated substances at the end of logarithmic growth in a semi-synthetic medium. Two of these substances are guanosine 3′(2′)-diphosphate 5′-diphosphate (ppGpp) and guanosine 3′(2′)-diphosphate 5′-triphosphate (pppGpp) which, in contrast to amino-acid-starved Escherichia coli cells, are not degraded in sporulating cells of B. subtilis after the addition of chloramphenicol. Moreover, inhibition of protein synthesis in growing cells of B. subtilis causes accumulation of ppGpp and pppGpp, which is also in contrast to E. coli. This was shown by isolation and characterization of substances produced in these cells after the addition of chloramphenicol. Other inhibitors of protein synthesis acting at the ribosomal level also cause the accumulation of ppGpp and pppGpp. There is no difference between the action of antibiotics affecting 50-S and/or 30-S ribosomal subunits, since chloramphenicol, tetracycline, erythromycin and neomycin cause the accumulation of almost equal amounts of these nucleotides. This apparently resolves the close connection between ppGpp accumulation and the rate of stable RNA synthesis, which was believed to exist also in B. subtilis because of the stringent response observed after amino acid starvation coupled with ppGpp accumulation. Antibiotics which inhibit protein synthesis differently than by affecting the ribosomes (puromycin) or which inhibit RNA (rifampicin) or DNA (nalidixic acid) synthesis do not cause ppGpp accumulation. The accumulation of ppGpp and pppGpp in the presence of charged tRNA provided by chloramphenicol treatment suggests that the signal for the synthesis of unusual nucleotides is an inhibition of the binding of tRNA (charged or uncharged) to the acceptor site of the ribosome. This activates the rel gene product which forms ppGpp and pppGpp from GTP and ATP. Sporulating cells of B. subtilis without chloramphenicol treatment produce besides ppGpp and pppGpp other unusual substances, which are likely to be highly phosphorylated nucleotides contained adenine as base moiety.Keywords
This publication has 17 references indexed in Scilit:
- Studies on the control of development. In vitro synthesis of HPN and MS nucleotides by ribosomes from either sporulating or vegetative cells of Bacillus subtilisFEBS Letters, 1974
- Inapparent correlation between guanosine tetraphosphate levels and RNA contents in Escherichia coliBiochemical and Biophysical Research Communications, 1974
- Metabolism of Guanosine Tetraphosphate in Escherichia coliEuropean Journal of Biochemistry, 1972
- Decoding site of initiator transfer RNABiochimie, 1971
- Inhibition of RNA Polymerase by ppGpp, a Nucleotide Accumulated during the Stringent Response to Amino Acid Starvation in E. coliCold Spring Harbor Symposia on Quantitative Biology, 1970
- An improved method for thin-layer chromatography of nucleotide mixtures containing32P-labeled orthophosphateJournal of Chromatography A, 1969
- Two Compounds implicated in the Function of the RC Gene of Escherichia coliNature, 1969
- Quantitative regulation of RNA synthesis during sporulation of bacillus subtilisBiochemical and Biophysical Research Communications, 1964
- Regulation of ribosomal and transfer RNA synthesisJournal of Molecular Biology, 1962
- Rôle des acides amines dans la synthèse des acides nucléiques chez Escherichia coliExperimental Cell Research, 1958