Summary
RNA polymerase was isolated from two suppressed rpoB amber strains exhibiting relaxed control over RNA synthesis in vivo. In vitro transcription analysis of these mutant holoenzymes carrying defined substitutions at known sites in the β subunit clearly shows that single amino acid changes in β render RNA polymerase resistant to ppGpp. This unambiguously demonstrates that RNA polymerase is indeed the target for ppGpp.
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References
Boothroyd C, Malet R, Nene V, Glass RE (1983) Genetic studies on the β subunit of Escherichia coli RNA polymerase, III: Analysis of low-level rifampicin resistant mutants. Mol Gen Genet 190:523–526
Burgess RR, Jendrisak JJ (1975) A procedure for the rapid large-scale purification of Escherichia coli DNA-dependent RNA polymerase involving polymin P precipitation and DNA-cellulose chromatography. Biochem 14:4634–4638
Friesen JD (1966) Control of messenger RNA synthesis and decay in Escherichia coli. J Mol Biol 20:559–573
Fukuda R, Iwakura Y, Ishihama A (1974) Heterogeneity of RNA polymerase in Escherichia coli I: A new holoenzyme containing a new sigma factor. J Mol Biol 83:353–367
Gallant JA (1979) Stringent control of E. coli Annu Rev Genet 13:393–415
Gonzalez N, Wiggs J, Chamberlin M (1977) A simple procedure for resolution of Escherichia coli RNA polymerase holoenzyme from core polymerase. Arch Biochem Biophys 182:404–408
Haseltine WA, Block R (1973) Synthesis of guanosine tetra- and pentaphosphate requires the presence of a codon specific, uncharged transfer ribonucleic acid in the acceptor site of ribosomes. Proc Natl Acad Sci USA 70:1564–1568
Kajitani M, Ishihama A (1983a) Determination of the promoter strength in the mixed transcription system: promoters of lactose, tryptophan and ribosomal protein L10 operons from Escherichia coli. Nucl Acids Res 11:671–686
Kajitani M, Ishihama A (1983b) Determination of the promoter strength in the mixed transcription system, II: Promoters of ribosomal RNA, ribosomal protein S1 and recA protein operons from Escherichia coli. Nucl Acids Res 11:3873–3889
Kajitani M, Ishihama A (1984) Promoter selectivity of Escherichia coli RNA polymerase: Differential stringent control of the multiple promoters from ribosomal RNA and protein operons. J Biol Chem 259:1951–1957
Lowe PA, Hager DA, Burgess RR (1979) Purification and properties of the σ subunit of Escherichia coli DNA-dependent RNA polymerase. Biochem 18:1344–1352
Nene V, Glass RE (1982) Genetic studies on the β subunit of Escherichia coli RNA polymerase, I: The effect of known, single amino acid substitutions in an essential protein. Mol Gen Genet 188:399–404
Nene V, Glass RE (1983) Relaxed mutants of Escherichia coli RNA polymerase. FEBS Lett 153:307–310
Nene V, Glass RE (1984) Genetic studies on the β subunit of Escherichia coli RNA polymerase, VI: A redundant region in the β polypeptide. Mol Gen Genet 196:64–67
Nunn WD, Cronan JE Jr (1976) Regulation of membrane phospholipid synthesis by the relA gene: Dependence on ppGpp levels. Biochem 15:2546–2550
Ovchinnikov YuA, Monastyrskaya GS, Gubanov VV, Guryev SO, Chertov OYu, Modyanov NN, Grinkevich UA, Makarova IA, Marchenko TV, Polovnikova IN, Lipkin VM, Sverdlov ED (1981) The primary structure of Escherichia coli RNA polymerase. Nucleotide sequence of the rpoB gene and amino acid sequence of the β subunit. Eur J Biochem 116:621–629
Pedersen FS, Lund E, Kejeldgaard NO (1973) Codon specific, tRNA dependent in vitro synthesis of ppGpp and pppGpp. Nature 234:13–15
Ryals J, Little R, Bremer H (1982) Control of rRNA and tRNA synthesis in Escherichia coli by guanosine tetraphosphate. J Bacteriol 151:1261–1268
Travers A (1976) Modulation of RNA polymerase specificity by ppGpp. Mol Gen Genet 147:225–232
Van Ooyen AJJ, Gruber M, Jorgensen P (1976) Mechanism of action of ppGpp on rRNA synthesis in vitro. Cell 8:123–128
Yang H-L, Zubay G, Urm E, Reiness G, Cashel M (1974) Effects of guanosine pentaphosphate and β-γ methyenyl-guanosine pentaphosphate on gene expression of Escherichia coli in vitro. Proc Natl Acad Sci USA 71:63–68
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Communicated by K. Isono
Paper VI is Nene and Glass (1984)
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Glass, R.E., Jones, S.T. & Ishihama, A. Genetic studies on the β subunit of Escherichia coli RNA polymerase. Molec Gen Genet 203, 265–268 (1986). https://doi.org/10.1007/BF00333964
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DOI: https://doi.org/10.1007/BF00333964