Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Jun;174(12):3903–3914. doi: 10.1128/jb.174.12.3903-3914.1992

Function of a relaxed-like state following temperature downshifts in Escherichia coli.

P G Jones 1, M Cashel 1, G Glaser 1, F C Neidhardt 1
PMCID: PMC206098  PMID: 1597413

Abstract

Temperature downshifts of Escherichia coli throughout its growth range resulted in transient growth inhibition and a cold shock response consisting of transient induction of several proteins, repression of heat shock proteins, and, despite the growth lag, continued synthesis of proteins involved in transcription and translation. The paradoxical synthesis of the latter proteins, which are normally repressed when growth is arrested, was explored further. First, by means of a nutritional downshift, a natural stringent response was induced in wild-type cells immediately prior to a shift from 37 to 10 degrees C. These cells displayed decreased synthesis of transcriptional and translational proteins and decreased induction of cold shock proteins; also, adaptation for growth at 10 degrees C was delayed, even after restoration of the nutrient supplementation. Next, the contribution of guanosine 5'-triphosphate-3'-diphosphate and guanosine 5'-diphosphate-3'-diphosphate, collectively abbreviated (p)ppGpp, to the alteration in cold shock response was studied with the aid of a mutant strain in which overproduction of these nucleotides can be artificially induced. Induction of (p)ppGpp synthesis immediately prior to shifting this strain from 37 to 10 degrees C produced results differing only in a few details from those described above for nutritional downshift of the wild-type strain. Finally, shifting a relA spoT mutant, which cannot synthesize (p)ppGpp, from 24 to 10 degrees C resulted in a greater induction of the cold shock proteins, increased synthesis of transcriptional and translational proteins, decreased synthesis of a major heat shock protein, and faster adaptation to growth than for the wild-type strain. Our results indicate that the previously reported decrease in the (p)ppGpp level following temperature downshift plays a physiological role in the regulation of gene expression and adaptation for growth at low temperature.

Full text

PDF
3903

Images in this article

3905Image
on p.3905
3905Image
on p.3905
3906Image
on p.3906
3906Image
on p.3906
3907Image
on p.3907
3909Image
on p.3909
3910Image
on p.3910
3911Image
on p.3911
3913Image
on p.3913

Selected References

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

  1. Broeze R. J., Solomon C. J., Pope D. H. Effects of low temperature on in vivo and in vitro protein synthesis in Escherichia coli and Pseudomonas fluorescens. J Bacteriol. 1978 Jun;134(3):861–874. doi: 10.1128/jb.134.3.861-874.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cortay J. C., Cozzone A. J. Effects of aminoglycoside antibiotics on the coupling of protein and RNA syntheses in Escherichia coli. Biochem Biophys Res Commun. 1983 May 16;112(3):801–808. doi: 10.1016/0006-291x(83)91688-1. [DOI] [PubMed] [Google Scholar]
  3. Didier D. K., Schiffenbauer J., Woulfe S. L., Zacheis M., Schwartz B. D. Characterization of the cDNA encoding a protein binding to the major histocompatibility complex class II Y box. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7322–7326. doi: 10.1073/pnas.85.19.7322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goldstein J., Pollitt N. S., Inouye M. Major cold shock protein of Escherichia coli. Proc Natl Acad Sci U S A. 1990 Jan;87(1):283–287. doi: 10.1073/pnas.87.1.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Granston A. E., Thompson D. L., Friedman D. I. Identification of a second promoter for the metY-nusA-infB operon of Escherichia coli. J Bacteriol. 1990 May;172(5):2336–2342. doi: 10.1128/jb.172.5.2336-2342.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Göransson M., Sondén B., Nilsson P., Dagberg B., Forsman K., Emanuelsson K., Uhlin B. E. Transcriptional silencing and thermoregulation of gene expression in Escherichia coli. Nature. 1990 Apr 12;344(6267):682–685. doi: 10.1038/344682a0. [DOI] [PubMed] [Google Scholar]
  7. Herendeen S. L., VanBogelen R. A., Neidhardt F. C. Levels of major proteins of Escherichia coli during growth at different temperatures. J Bacteriol. 1979 Jul;139(1):185–194. doi: 10.1128/jb.139.1.185-194.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hernandez V. J., Bremer H. Escherichia coli ppGpp synthetase II activity requires spoT. J Biol Chem. 1991 Mar 25;266(9):5991–5999. [PubMed] [Google Scholar]
  9. Jones P. G., VanBogelen R. A., Neidhardt F. C. Induction of proteins in response to low temperature in Escherichia coli. J Bacteriol. 1987 May;169(5):2092–2095. doi: 10.1128/jb.169.5.2092-2095.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. La Teana A., Brandi A., Falconi M., Spurio R., Pon C. L., Gualerzi C. O. Identification of a cold shock transcriptional enhancer of the Escherichia coli gene encoding nucleoid protein H-NS. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10907–10911. doi: 10.1073/pnas.88.23.10907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mackow E. R., Chang F. N. Correlation between RNA synthesis and ppGpp content in Escherichia coli during temperature shifts. Mol Gen Genet. 1983;192(1-2):5–9. doi: 10.1007/BF00327639. [DOI] [PubMed] [Google Scholar]
  12. Metzger S., Sarubbi E., Glaser G., Cashel M. Protein sequences encoded by the relA and the spoT genes of Escherichia coli are interrelated. J Biol Chem. 1989 Jun 5;264(16):9122–9125. [PubMed] [Google Scholar]
  13. Metzger S., Schreiber G., Aizenman E., Cashel M., Glaser G. Characterization of the relA1 mutation and a comparison of relA1 with new relA null alleles in Escherichia coli. J Biol Chem. 1989 Dec 15;264(35):21146–21152. [PubMed] [Google Scholar]
  14. NG H., INGRAHAM J. L., MARR A. G. Damage and derepression in Escherichia coli resulting from growth at low temperatures. J Bacteriol. 1962 Aug;84:331–339. doi: 10.1128/jb.84.2.331-339.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  17. Pao C. C., Dyess B. T. Stringent control of RNA synthesis in the absence of guanosine 5'-diphosphate-3'-diphosphate. J Biol Chem. 1981 Mar 10;256(5):2252–2257. [PubMed] [Google Scholar]
  18. Rojiani M. V., Jakubowski H., Goldman E. Effect of variation of charged and uncharged tRNA(Trp) levels on ppGpp synthesis in Escherichia coli. J Bacteriol. 1989 Dec;171(12):6493–6502. doi: 10.1128/jb.171.12.6493-6502.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rojiani M. V., Jakubowski H., Goldman E. Relationship between protein synthesis and concentrations of charged and uncharged tRNATrp in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1511–1515. doi: 10.1073/pnas.87.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Sakura H., Maekawa T., Imamoto F., Yasuda K., Ishii S. Two human genes isolated by a novel method encode DNA-binding proteins containing a common region of homology. Gene. 1988 Dec 20;73(2):499–507. doi: 10.1016/0378-1119(88)90514-8. [DOI] [PubMed] [Google Scholar]
  23. Schreiber G., Metzger S., Aizenman E., Roza S., Cashel M., Glaser G. Overexpression of the relA gene in Escherichia coli. J Biol Chem. 1991 Feb 25;266(6):3760–3767. [PubMed] [Google Scholar]
  24. Tafuri S. R., Wolffe A. P. Xenopus Y-box transcription factors: molecular cloning, functional analysis and developmental regulation. Proc Natl Acad Sci U S A. 1990 Nov;87(22):9028–9032. doi: 10.1073/pnas.87.22.9028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Takata R., Mukai T., Hori K. Attenuation and processing of RNA from the rpsO-pnp transcription unit of Escherichia coli. Nucleic Acids Res. 1985 Oct 25;13(20):7289–7297. doi: 10.1093/nar/13.20.7289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Taura T., Kusukawa N., Yura T., Ito K. Transient shut off of Escherichia coli heat shock protein synthesis upon temperature shift down. Biochem Biophys Res Commun. 1989 Aug 30;163(1):438–443. doi: 10.1016/0006-291x(89)92155-4. [DOI] [PubMed] [Google Scholar]
  27. VanBogelen R. A., Neidhardt F. C. Ribosomes as sensors of heat and cold shock in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5589–5593. doi: 10.1073/pnas.87.15.5589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wanner B. L., Kodaira R., Neidhardt F. C. Physiological regulation of a decontrolled lac operon. J Bacteriol. 1977 Apr;130(1):212–222. doi: 10.1128/jb.130.1.212-222.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wistow G. Cold shock and DNA binding. Nature. 1990 Apr 26;344(6269):823–824. doi: 10.1038/344823c0. [DOI] [PubMed] [Google Scholar]
  30. Xiao H., Kalman M., Ikehara K., Zemel S., Glaser G., Cashel M. Residual guanosine 3',5'-bispyrophosphate synthetic activity of relA null mutants can be eliminated by spoT null mutations. J Biol Chem. 1991 Mar 25;266(9):5980–5990. [PubMed] [Google Scholar]

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

RESOURCES