Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Nov;179(22):7081–7088. doi: 10.1128/jb.179.22.7081-7088.1997

CspA, the major cold shock protein of Escherichia coli, negatively regulates its own gene expression.

W Bae 1, P G Jones 1, M Inouye 1
PMCID: PMC179650  PMID: 9371456

Abstract

When the gene for CspA, the major cold shock protein of Escherichia coli, was disrupted by a novel positive/negative selection method, the deltacspA cells did not show any discernible growth defect at either 37 or 15 degrees C. By two-dimensional gel electrophoresis, total protein synthesis was analyzed after temperature downshift in the deltacspA strain. The production of the CspA homologs CspB and CspG increased, and the duration of their expression was prolonged, suggesting that both CspB and CspG compensate for the function of CspA in the absence of CspA during cold shock adaptation. Interestingly, the production of the 159-base 5'-untranslated region (5'-UTR) of cspA from the chromosomal cspA::cat gene, detected by primer extension, failed to be repressed after cold shock. When an independent system to produce CspA was added to the deltacspA strain, the 5'-UTR production for the cspA::cat gene was significantly reduced compared to that of the deltacspA strain. By examining the expression of translationally fused cspA and cspB genes to lacZ in the deltacspA strain, it was found that cspA is more strongly regulated by CspA than cspB is. We showed that the increased expression of the 5'-UTR of the cspA mRNA in the deltacspA strain occurred mainly at the level of transcription and, to a certain extent, at the level of mRNA stabilization. The mRNA stabilization in the deltacspA strain was observed for other mRNAs, supporting the notion that CspA functions as an mRNA chaperone to destabilize secondary structures in mRNAs.

Full Text

The Full Text of this article is available as a PDF (505.4 KB).

Selected References

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

  1. Brandi A., Pietroni P., Gualerzi C. O., Pon C. L. Post-transcriptional regulation of CspA expression in Escherichia coli. Mol Microbiol. 1996 Jan;19(2):231–240. doi: 10.1046/j.1365-2958.1996.362897.x. [DOI] [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Das A. Control of transcription termination by RNA-binding proteins. Annu Rev Biochem. 1993;62:893–930. doi: 10.1146/annurev.bi.62.070193.004333. [DOI] [PubMed] [Google Scholar]
  4. Etchegaray J. P., Jones P. G., Inouye M. Differential thermoregulation of two highly homologous cold-shock genes, cspA and cspB, of Escherichia coli. Genes Cells. 1996 Feb;1(2):171–178. doi: 10.1046/j.1365-2443.1996.d01-231.x. [DOI] [PubMed] [Google Scholar]
  5. Goldenberg D., Azar I., Oppenheim A. B. Differential mRNA stability of the cspA gene in the cold-shock response of Escherichia coli. Mol Microbiol. 1996 Jan;19(2):241–248. doi: 10.1046/j.1365-2958.1996.363898.x. [DOI] [PubMed] [Google Scholar]
  6. Gottesman S., Clark W. P., Maurizi M. R. The ATP-dependent Clp protease of Escherichia coli. Sequence of clpA and identification of a Clp-specific substrate. J Biol Chem. 1990 May 15;265(14):7886–7893. [PubMed] [Google Scholar]
  7. Harlocker S. L., Rampersaud A., Yang W. P., Inouye M. Phenotypic revertant mutations of a new OmpR2 mutant (V203Q) of Escherichia coli lie in the envZ gene, which encodes the OmpR kinase. J Bacteriol. 1993 Apr;175(7):1956–1960. doi: 10.1128/jb.175.7.1956-1960.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jiang W., Fang L., Inouye M. Complete growth inhibition of Escherichia coli by ribosome trapping with truncated cspA mRNA at low temperature. Genes Cells. 1996 Nov;1(11):965–976. doi: 10.1046/j.1365-2443.1996.d01-219.x. [DOI] [PubMed] [Google Scholar]
  9. Jiang W., Fang L., Inouye M. The role of the 5'-end untranslated region of the mRNA for CspA, the major cold-shock protein of Escherichia coli, in cold-shock adaptation. J Bacteriol. 1996 Aug;178(16):4919–4925. doi: 10.1128/jb.178.16.4919-4925.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jiang W., Hou Y., Inouye M. CspA, the major cold-shock protein of Escherichia coli, is an RNA chaperone. J Biol Chem. 1997 Jan 3;272(1):196–202. doi: 10.1074/jbc.272.1.196. [DOI] [PubMed] [Google Scholar]
  11. Jiang W., Jones P., Inouye M. Chloramphenicol induces the transcription of the major cold shock gene of Escherichia coli, cspA. J Bacteriol. 1993 Sep;175(18):5824–5828. doi: 10.1128/jb.175.18.5824-5828.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jones P. G., Inouye M. RbfA, a 30S ribosomal binding factor, is a cold-shock protein whose absence triggers the cold-shock response. Mol Microbiol. 1996 Sep;21(6):1207–1218. doi: 10.1111/j.1365-2958.1996.tb02582.x. [DOI] [PubMed] [Google Scholar]
  13. Jones P. G., Inouye M. The cold-shock response--a hot topic. Mol Microbiol. 1994 Mar;11(5):811–818. doi: 10.1111/j.1365-2958.1994.tb00359.x. [DOI] [PubMed] [Google Scholar]
  14. Jones P. G., Mitta M., Kim Y., Jiang W., Inouye M. Cold shock induces a major ribosomal-associated protein that unwinds double-stranded RNA in Escherichia coli. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):76–80. doi: 10.1073/pnas.93.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Lee S. J., Xie A., Jiang W., Etchegaray J. P., Jones P. G., Inouye M. Family of the major cold-shock protein, CspA (CS7.4), of Escherichia coli, whose members show a high sequence similarity with the eukaryotic Y-box binding proteins. Mol Microbiol. 1994 Mar;11(5):833–839. doi: 10.1111/j.1365-2958.1994.tb00361.x. [DOI] [PubMed] [Google Scholar]
  17. Lloyd R. G., Buckman C. Identification and genetic analysis of sbcC mutations in commonly used recBC sbcB strains of Escherichia coli K-12. J Bacteriol. 1985 Nov;164(2):836–844. doi: 10.1128/jb.164.2.836-844.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nakashima K., Kanamaru K., Mizuno T., Horikoshi K. A novel member of the cspA family of genes that is induced by cold shock in Escherichia coli. J Bacteriol. 1996 May;178(10):2994–2997. doi: 10.1128/jb.178.10.2994-2997.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Newkirk K., Feng W., Jiang W., Tejero R., Emerson S. D., Inouye M., Montelione G. T. Solution NMR structure of the major cold shock protein (CspA) from Escherichia coli: identification of a binding epitope for DNA. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5114–5118. doi: 10.1073/pnas.91.11.5114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., Inada T., Itoh T., Kajihara M. A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 12.7-28.0 min region on the linkage map. DNA Res. 1996 Jun 30;3(3):137–155. doi: 10.1093/dnares/3.3.137. [DOI] [PubMed] [Google Scholar]
  21. Platt T. Transcription termination and the regulation of gene expression. Annu Rev Biochem. 1986;55:339–372. doi: 10.1146/annurev.bi.55.070186.002011. [DOI] [PubMed] [Google Scholar]
  22. Schindelin H., Jiang W., Inouye M., Heinemann U. Crystal structure of CspA, the major cold shock protein of Escherichia coli. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5119–5123. doi: 10.1073/pnas.91.11.5119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sprengart M. L., Fuchs E., Porter A. G. The downstream box: an efficient and independent translation initiation signal in Escherichia coli. EMBO J. 1996 Feb 1;15(3):665–674. [PMC free article] [PubMed] [Google Scholar]
  24. Tanabe H., Goldstein J., Yang M., Inouye M. Identification of the promoter region of the Escherichia coli major cold shock gene, cspA. J Bacteriol. 1992 Jun;174(12):3867–3873. doi: 10.1128/jb.174.12.3867-3873.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ueki T., Inouye S., Inouye M. Positive-negative KG cassettes for construction of multi-gene deletions using a single drug marker. Gene. 1996 Dec 12;183(1-2):153–157. doi: 10.1016/s0378-1119(96)00546-x. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Wolffe A. P. Structural and functional properties of the evolutionarily ancient Y-box family of nucleic acid binding proteins. Bioessays. 1994 Apr;16(4):245–251. doi: 10.1002/bies.950160407. [DOI] [PubMed] [Google Scholar]
  28. Wu C. J., Janssen G. R. Translation of vph mRNA in Streptomyces lividans and Escherichia coli after removal of the 5' untranslated leader. Mol Microbiol. 1996 Oct;22(2):339–355. doi: 10.1046/j.1365-2958.1996.00119.x. [DOI] [PubMed] [Google Scholar]
  29. Yamanaka K., Mitani T., Ogura T., Niki H., Hiraga S. Cloning, sequencing, and characterization of multicopy suppressors of a mukB mutation in Escherichia coli. Mol Microbiol. 1994 Jul;13(2):301–312. doi: 10.1111/j.1365-2958.1994.tb00424.x. [DOI] [PubMed] [Google Scholar]
  30. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES