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. 1989 Mar;171(3):1712–1717. doi: 10.1128/jb.171.3.1712-1717.1989

Suppression of the Escherichia coli rpoH opal mutation by ribosomes lacking S15 protein.

R Yano 1, T Yura 1
PMCID: PMC209802  PMID: 2646293

Abstract

Several suppressors (suhD) that can specifically suppress the temperature-sensitive opal rpoH11 mutation of Escherichia coli K-12 have been isolated and characterized. Unlike the parental rpoH11 mutant deficient in the heat shock response, the temperature-resistant pseudorevertants carrying suhD were capable of synthesizing sigma 32 and exhibiting partial induction of heat shock proteins. These strains were also cold sensitive and unable to grow at 25 degrees C. Genetic mapping and complementation studies permitted us to localize suhD near rpsO (69 min), the structural gene for ribosomal protein S15. Ribosomes and polyribosomes prepared from suhD cells contained a reduced level (ca. 10%) of S15 relative to that of the wild type. Cloning and sequencing of suhD revealed that an IS10-like element had been inserted at the attenuator-terminator region immediately downstream of the rpsO coding region. The rpsO mRNA level in the suhD strain was also reduced to about 10% that of wild type. Apparently, ribosomes lacking S15 can actively participate in protein synthesis and suppress the rpoH11 opal (UGA) mutation at high temperature but cannot sustain cell growth at low temperature.

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Selected References

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  1. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  2. Cole J. R., Olsson C. L., Hershey J. W., Grunberg-Manago M., Nomura M. Feedback regulation of rRNA synthesis in Escherichia coli. Requirement for initiation factor IF2. J Mol Biol. 1987 Dec 5;198(3):383–392. doi: 10.1016/0022-2836(87)90288-9. [DOI] [PubMed] [Google Scholar]
  3. Dabbs E. R., Hasenbank R., Kastner B., Rak K. H., Wartusch B., Stöffler G. Immunological studies of Escherichia coli mutants lacking one or two ribosomal proteins. Mol Gen Genet. 1983;192(3):301–308. doi: 10.1007/BF00392166. [DOI] [PubMed] [Google Scholar]
  4. Geyl D., Böck A., Isono K. An improved method for two-dimensional gel-electrophoresis: analysis of mutationally altered ribosomal proteins of Escherichia coli. Mol Gen Genet. 1981;181(3):309–312. doi: 10.1007/BF00425603. [DOI] [PubMed] [Google Scholar]
  5. Grossman A. D., Erickson J. W., Gross C. A. The htpR gene product of E. coli is a sigma factor for heat-shock promoters. Cell. 1984 Sep;38(2):383–390. doi: 10.1016/0092-8674(84)90493-8. [DOI] [PubMed] [Google Scholar]
  6. Halling S. M., Simons R. W., Way J. C., Walsh R. B., Kleckner N. DNA sequence organization of IS10-right of Tn10 and comparison with IS10-left. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2608–2612. doi: 10.1073/pnas.79.8.2608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Held W. A., Ballou B., Mizushima S., Nomura M. Assembly mapping of 30 S ribosomal proteins from Escherichia coli. Further studies. J Biol Chem. 1974 May 25;249(10):3103–3111. [PubMed] [Google Scholar]
  8. Ito K., Cerretti D. P., Nashimoto H., Nomura M. Characterization of an amber mutation in the structural gene for ribosomal protein L15, which impairs the expression of the protein export gene, secY, in Escherichia coli. EMBO J. 1984 Oct;3(10):2319–2324. doi: 10.1002/j.1460-2075.1984.tb02133.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kirsebom L. A., Isaksson L. A. Involvement of ribosomal protein L7/L12 in control of translational accuracy. Proc Natl Acad Sci U S A. 1985 Feb;82(3):717–721. doi: 10.1073/pnas.82.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kusukawa N., Yura T. Heat shock protein GroE of Escherichia coli: key protective roles against thermal stress. Genes Dev. 1988 Jul;2(7):874–882. doi: 10.1101/gad.2.7.874. [DOI] [PubMed] [Google Scholar]
  11. Lemaux P. G., Herendeen S. L., Bloch P. L., Neidhardt F. C. Transient rates of synthesis of individual polypeptides in E. coli following temperature shifts. Cell. 1978 Mar;13(3):427–434. doi: 10.1016/0092-8674(78)90317-3. [DOI] [PubMed] [Google Scholar]
  12. Mackie G. A. Posttranscriptional regulation of ribosomal protein S20 and stability of the S20 mRNA species. J Bacteriol. 1987 Jun;169(6):2697–2701. doi: 10.1128/jb.169.6.2697-2701.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mizusawa S., Nishimura S., Seela F. Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP. Nucleic Acids Res. 1986 Feb 11;14(3):1319–1324. doi: 10.1093/nar/14.3.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mori K., Aiba H. Evidence for negative control of cya transcription by cAMP and cAMP receptor protein in intact Escherichia coli cells. J Biol Chem. 1985 Nov 25;260(27):14838–14843. [PubMed] [Google Scholar]
  15. Murgola E. J., Hijazi K. A., Göringer H. U., Dahlberg A. E. Mutant 16S ribosomal RNA: a codon-specific translational suppressor. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4162–4165. doi: 10.1073/pnas.85.12.4162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nakamura Y., Mizusawa S. In vivo evidence that the nusA and infB genes of E. coli are part of the same multi-gene operon which encodes at least four proteins. EMBO J. 1985 Feb;4(2):527–532. doi: 10.1002/j.1460-2075.1985.tb03660.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nashimoto H., Nomura M. Structure and function of bacterial ribosomes. XI. Dependence of 50S ribosomal assembly on simultaneous assembly of 30S subunits. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1440–1447. doi: 10.1073/pnas.67.3.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Neidhardt F. C., VanBogelen R. A. Positive regulatory gene for temperature-controlled proteins in Escherichia coli. Biochem Biophys Res Commun. 1981 May 29;100(2):894–900. doi: 10.1016/s0006-291x(81)80257-4. [DOI] [PubMed] [Google Scholar]
  19. Portier C., Regnier P. Expression of the rpsO and pnp genes: structural analysis of a DNA fragment carrying their control regions. Nucleic Acids Res. 1984 Aug 10;12(15):6091–6102. doi: 10.1093/nar/12.15.6091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Régnier P., Portier C. Initiation, attenuation and RNase III processing of transcripts from the Escherichia coli operon encoding ribosomal protein S15 and polynucleotide phosphorylase. J Mol Biol. 1986 Jan 5;187(1):23–32. doi: 10.1016/0022-2836(86)90403-1. [DOI] [PubMed] [Google Scholar]
  21. Shiba K., Ito K., Yura T. Suppressors of the secY24 mutation: identification and characterization of additional ssy genes in Escherichia coli. J Bacteriol. 1986 Jun;166(3):849–856. doi: 10.1128/jb.166.3.849-856.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Straus D. B., Walter W. A., Gross C. A. The heat shock response of E. coli is regulated by changes in the concentration of sigma 32. Nature. 1987 Sep 24;329(6137):348–351. doi: 10.1038/329348a0. [DOI] [PubMed] [Google Scholar]
  23. Svensson P., Changchien L. M., Craven G. R., Noller H. F. Interaction of ribosomal proteins, S6, S8, S15 and S18 with the central domain of 16 S ribosomal RNA. J Mol Biol. 1988 Mar 20;200(2):301–308. doi: 10.1016/0022-2836(88)90242-2. [DOI] [PubMed] [Google Scholar]
  24. Takata R. Genetic studies of the ribosomal proteins in Escherichia coli. XI. Mapping of the genes for L21, L27, S15 and S21 by using hybrid bacteria and over-production of these proteins in the merodiploid strains. Mol Gen Genet. 1978 Apr 6;160(2):151–155. doi: 10.1007/BF00267476. [DOI] [PubMed] [Google Scholar]
  25. Takata R., Mukai T., Aoyagi M., Hori K. Nucleotide sequence of the gene for Escherichia coli ribosomal protein S15 (rpsO). Mol Gen Genet. 1984;197(2):225–229. doi: 10.1007/BF00330967. [DOI] [PubMed] [Google Scholar]
  26. Tobe T., Ito K., Yura T. Isolation and physical mapping of temperature-sensitive mutants defective in heat-shock induction of proteins in Escherichia coli. Mol Gen Genet. 1984;195(1-2):10–16. doi: 10.1007/BF00332716. [DOI] [PubMed] [Google Scholar]
  27. Tobe T., Kusukawa N., Yura T. Suppression of rpoH (htpR) mutations of Escherichia coli: heat shock response in suhA revertants. J Bacteriol. 1987 Sep;169(9):4128–4134. doi: 10.1128/jb.169.9.4128-4134.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  29. Vijgenboom E., Vink T., Kraal B., Bosch L. Mutants of the elongation factor EF-Tu, a new class of nonsense suppressors. EMBO J. 1985 Apr;4(4):1049–1052. doi: 10.1002/j.1460-2075.1985.tb03737.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yamamori T., Ito K., Nakamura Y., Yura T. Transient regulation of protein synthesis in Escherichia coli upon shift-up of growth temperature. J Bacteriol. 1978 Jun;134(3):1133–1140. doi: 10.1128/jb.134.3.1133-1140.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yamamori T., Yura T. Genetic control of heat-shock protein synthesis and its bearing on growth and thermal resistance in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1982 Feb;79(3):860–864. doi: 10.1073/pnas.79.3.860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yamamori T., Yura T. Temperature-induced synthesis of specific proteins in Escherichia coli: evidence for transcriptional control. J Bacteriol. 1980 Jun;142(3):843–851. doi: 10.1128/jb.142.3.843-851.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yano R., Imai M., Yura T. The use of operon fusions in studies of the heat-shock response: effects of altered sigma 32 on heat-shock promoter function in Escherichia coli. Mol Gen Genet. 1987 Apr;207(1):24–28. doi: 10.1007/BF00331486. [DOI] [PubMed] [Google Scholar]
  34. Yura T., Tobe T., Ito K., Osawa T. Heat shock regulatory gene (htpR) of Escherichia coli is required for growth at high temperature but is dispensable at low temperature. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6803–6807. doi: 10.1073/pnas.81.21.6803. [DOI] [PMC free article] [PubMed] [Google Scholar]

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