Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Sep;155(3):1162–1170. doi: 10.1128/jb.155.3.1162-1170.1983

Physiological characterization of Escherichia coli rpoB mutants with abnormal control of ribosome synthesis.

R Little, J Ryals, H Bremer
PMCID: PMC217812  PMID: 6193095

Abstract

We have previously reported the isolation of Escherichia coli rpoB mutants in which the control of ribosome synthesis by the nucleotide effector guanosine tetraphosphate (ppGpp) is altered, owing to a 20-fold increased sensitivity of the mutant RNA polymerases to ppGpp. In these mutants, the level of ppGpp during exponential growth is decreased about 10-fold, relative to that of rpoB+ wild-type strains, such that a near normal partitioning of RNA polymerase occurs with respect to stable RNA (rRNA and tRNA) gene activity. Here, the physiological effects of two different rpoB alleles in a relA+ and relA background were analyzed in greater detail by comparison with their isogenic rpoB+ wild-type parents. For a given growth medium, the rpoB mutations were found to affect four parameters which resulted in a reduction of growth rate. The results reinforce a previous conclusion that a key element in control of the bacterial growth rate is a mutual relationship between control of ribosome synthesis by ppGpp and control of relA-independent ppGpp metabolism by the concentration and function of ribosomes.

Full text

PDF
1162

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barry G., Squires C. L., Squires C. Control features within the rplJL-rpoBC transcription unit of Escherichia coli. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4922–4926. doi: 10.1073/pnas.76.10.4922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blumenthal R. M., Dennis P. P. Gene expression in Escherichia coli after amino acid, purine, or pyrimidine exhaustion. J Bacteriol. 1980 Apr;142(1):202–211. doi: 10.1128/jb.142.1.202-211.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bremer H., Berry L., Dennis P. P. Regulation of ribonucleic acid synthesis in Escherichia coli B-r: an analysis of a shift-up. II. Fraction of RNA polymerase engaged in the synthesis of stable RNA at different steady-state growth rates. J Mol Biol. 1973 Mar 25;75(1):161–179. doi: 10.1016/0022-2836(73)90536-6. [DOI] [PubMed] [Google Scholar]
  4. Bremer H., Hymes J., Dennis P. P. Ribosomal RNA chain growth rate and RNA labeling patterns in Escherichia coli B-r. J Theor Biol. 1974 Jun;45(2):379–403. doi: 10.1016/0022-5193(74)90120-9. [DOI] [PubMed] [Google Scholar]
  5. Bremer H. Parameters affecting the rate of synthesis of ribosomes and RNA polymerase in bacteria. J Theor Biol. 1975 Sep;53(1):115–124. doi: 10.1016/0022-5193(75)90106-x. [DOI] [PubMed] [Google Scholar]
  6. Brunschede H., Dove T. L., Bremer H. Establishment of exponential growth after a nutritional shift-up in Escherichia coli B/r: accumulation of deoxyribonucleic acid, ribonucleic acid, and protein. J Bacteriol. 1977 Feb;129(2):1020–1033. doi: 10.1128/jb.129.2.1020-1033.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Churchward G., Bremer H., Young R. Macromolecular composition of bacteria. J Theor Biol. 1982 Feb 7;94(3):651–670. doi: 10.1016/0022-5193(82)90305-8. [DOI] [PubMed] [Google Scholar]
  8. Churchward G., Bremer H., Young R. Transcription in bacteria at different DNA concentrations. J Bacteriol. 1982 May;150(2):572–581. doi: 10.1128/jb.150.2.572-581.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dalbow D. G., Bremer H. Metabolic regulation of beta-galactosidase synthesis in Escherichia coli. A test for constitutive ribosome synthesis. Biochem J. 1975 Jul;150(1):1–8. doi: 10.1042/bj1500001b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dalbow D. G., Young R. Synthesis time of beta-galactosidase in Escherichia coli B/r as a function of growth rate. Biochem J. 1975 Jul;150(1):13–20. doi: 10.1042/bj1500013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dennis P. P., Bremer H. A method for determination of the synthesis rate of stable and unstable ribonucleic acid in Escherichia coli. Anal Biochem. 1973 Dec;56(2):489–501. doi: 10.1016/0003-2697(73)90216-9. [DOI] [PubMed] [Google Scholar]
  12. Dennis P. P. Regulation of synthesis and activity of a mutant RNA polymerase in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5416–5420. doi: 10.1073/pnas.74.12.5416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dennis P. P. Transcription patterns of adjacent segments on the chromosome of Escherichia coli containing genes coding for four 50S ribosomal proteins and the beta and beta' subunits of RNA polymerase. J Mol Biol. 1977 Oct 5;115(4):603–625. doi: 10.1016/0022-2836(77)90105-x. [DOI] [PubMed] [Google Scholar]
  14. Gausing K. Regulation of ribosome production in Escherichia coli: synthesis and stability of ribosomal RNA and of ribosomal protein messenger RNA at different growth rates. J Mol Biol. 1977 Sep 25;115(3):335–354. doi: 10.1016/0022-2836(77)90158-9. [DOI] [PubMed] [Google Scholar]
  15. Haseltine W. A., Block R., Gilbert W., Weber K. MSI and MSII made on ribosome in idling step of protein synthesis. Nature. 1972 Aug 18;238(5364):381–384. doi: 10.1038/238381a0. [DOI] [PubMed] [Google Scholar]
  16. Hui I., Maltman K., Little R., Hastrup S., Johnsen M., Fiil N., Dennis P. Insertions of transposon Tn5 into ribosomal protein PNA polymerase operons. J Bacteriol. 1982 Dec;152(3):1022–1032. doi: 10.1128/jb.152.3.1022-1032.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kingston R. E., Chamberlin M. J. Pausing and attenuation of in vitro transcription in the rrnB operon of E. coli. Cell. 1981 Dec;27(3 Pt 2):523–531. doi: 10.1016/0092-8674(81)90394-9. [DOI] [PubMed] [Google Scholar]
  18. Kingston R. E., Nierman W. C., Chamberlin M. J. A direct effect of guanosine tetraphosphate on pausing of Escherichia coli RNA polymerase during RNA chain elongation. J Biol Chem. 1981 Mar 25;256(6):2787–2797. [PubMed] [Google Scholar]
  19. Lindahl L., Zengel J. M. Expression of ribosomal genes in bacteria. Adv Genet. 1982;21:53–121. doi: 10.1016/s0065-2660(08)60297-7. [DOI] [PubMed] [Google Scholar]
  20. Linn T., Scaife J. Identification of a single promoter in E. coli for rplJ, rplL and rpoBC. Nature. 1978 Nov 2;276(5683):33–37. doi: 10.1038/276033a0. [DOI] [PubMed] [Google Scholar]
  21. Little R., Bremer H. Quantitation of guanosine 5',3'-bisdiphosphate in extracts from bacterial cells by ion-pair reverse-phase high-performance liquid chromatography. Anal Biochem. 1982 Nov 1;126(2):381–388. doi: 10.1016/0003-2697(82)90531-0. [DOI] [PubMed] [Google Scholar]
  22. Little R., Dennis P. P. Expression of RNA polymerase and ribosome component genes in Escherichia coli mutants having conditionally defective RNA polymerases. J Bacteriol. 1979 Jan;137(1):115–123. doi: 10.1128/jb.137.1.115-123.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Little R., Fiil N. P., Dennis P. P. Transcriptional and post-transcriptional control of ribosomal protein and ribonucleic acid polymerase genes. J Bacteriol. 1981 Jul;147(1):25–35. doi: 10.1128/jb.147.1.25-35.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Little R., Ryals J., Bremer H. rpoB mutation in Escherichia coli alters control of ribosome synthesis by guanosine tetraphosphate. J Bacteriol. 1983 May;154(2):787–792. doi: 10.1128/jb.154.2.787-792.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maher D. L., Dennis P. P. In vivo transcription of E. coli genes coding for rRNA, ribosomal proteins and subunits of RNA polymerase: influence of the stringent control system. Mol Gen Genet. 1977 Oct 20;155(2):203–211. doi: 10.1007/BF00393161. [DOI] [PubMed] [Google Scholar]
  26. Norris T. E., Koch A. L. Effect of growth rate on the relative rates of synthesis of messenger, ribosomal and transfer RNA in Escherichia coli. J Mol Biol. 1972 Mar 14;64(3):633–649. doi: 10.1016/0022-2836(72)90088-5. [DOI] [PubMed] [Google Scholar]
  27. Post L. E., Strycharz G. D., Nomura M., Lewis H., Dennis P. P. Nucleotide sequence of the ribosomal protein gene cluster adjacent to the gene for RNA polymerase subunit beta in Escherichia coli. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1697–1701. doi: 10.1073/pnas.76.4.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reeh S., Pedersen S., Friesen J. D. Biosynthetic regulation of individual proteins in relA+ and relA strains of Escherichia coli during amino acid starvation. Mol Gen Genet. 1976 Dec 22;149(3):279–289. doi: 10.1007/BF00268529. [DOI] [PubMed] [Google Scholar]
  29. Ryals J., Bremer H. relA-dependent RNA polymerase activity in Escherichia coli. J Bacteriol. 1982 Apr;150(1):168–179. doi: 10.1128/jb.150.1.168-179.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ryals J., Little R., Bremer H. Control of RNA synthesis in Escherichia coli after a shift to higher temperature. J Bacteriol. 1982 Sep;151(3):1425–1432. doi: 10.1128/jb.151.3.1425-1432.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ryals J., Little R., Bremer H. Control of rRNA and tRNA syntheses in Escherichia coli by guanosine tetraphosphate. J Bacteriol. 1982 Sep;151(3):1261–1268. doi: 10.1128/jb.151.3.1261-1268.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ryals J., Little R., Bremer H. Temperature dependence of RNA synthesis parameters in Escherichia coli. J Bacteriol. 1982 Aug;151(2):879–887. doi: 10.1128/jb.151.2.879-887.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shepherd N. S., Churchward G., Bremer H. Synthesis and activity of ribonucleic acid polymerase in Escherichia coli. J Bacteriol. 1980 Mar;141(3):1098–1108. doi: 10.1128/jb.141.3.1098-1108.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shepherd N., Churchward G., Bremer H. Synthesis and function of ribonucleic acid polymerase and ribosomes in Escherichia coli B/r after a nutritional shift-up. J Bacteriol. 1980 Sep;143(3):1332–1344. doi: 10.1128/jb.143.3.1332-1344.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Travers A. Modulation of RNA polymerase specificity by ppGpp. Mol Gen Genet. 1976 Aug 19;147(2):225–232. doi: 10.1007/BF00267575. [DOI] [PubMed] [Google Scholar]
  36. Yamamoto M., Nomura M. Contranscription of genes for RNA polymerase subunits beta and beta' with genes for ribosomal proteins in Escherichia coli. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3891–3895. doi: 10.1073/pnas.75.8.3891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Young R., Bremer H. Analysis of enzyme induction in bacteria. Biochem J. 1975 Nov;152(2):243–254. doi: 10.1042/bj1520243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. van Ooyen A. J., Gruber M., Jorgensen P. The mechanism of action of ppGpp on rRNA synthesis in vitro. Cell. 1976 May;8(1):123–128. doi: 10.1016/0092-8674(76)90193-8. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES