Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 May;171(5):2639–2645. doi: 10.1128/jb.171.5.2639-2645.1989

Translational coupling of the two proximal genes in the S10 ribosomal protein operon of Escherichia coli.

L Lindahl 1, R H Archer 1, J R McCormick 1, L P Freedman 1, J M Zengel 1
PMCID: PMC209946  PMID: 2651412

Abstract

We have examined the translational coupling between the first two genes in the S10 ribosomal protein operon. We isolated mutations blocking the translation of the first gene of the operon, coding for S10, and monitored their effects on translation of the downstream gene, coding for L3. All of the mutations inhibiting S10 synthesis also affected the synthesis of L3. However, these experiments were complicated by decreased mRNA synthesis resulting from transcription polarity, which we could only partially eliminate by using a rho-100 strain. To completely eliminate the problem of transcription polarity and obtain a more accurate measurement of the coupling, we replaced the natural S10 promoter with a promoter used by the bacteriophage T7 RNA polymerase. As expected, the T7 RNA polymerase was not subject to transcription polarity. Using this system, we were able to show that a complete abolishment of S10 translation resulted in an 80% inhibition of L3 synthesis. Other experiments show that the synthesis of L3 goes up as a function of increasing S10 synthesis, but the translational coupling does not assure strictly proportional output from the two genes.

Full text

PDF
2639

Images in this article

2642Image
on p.2642
2643Image
on p.2643

Selected References

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

  1. Adhya S., Gottesman M. Control of transcription termination. Annu Rev Biochem. 1978;47:967–996. doi: 10.1146/annurev.bi.47.070178.004535. [DOI] [PubMed] [Google Scholar]
  2. Aksoy S., Squires C. L., Squires C. Translational coupling of the trpB and trpA genes in the Escherichia coli tryptophan operon. J Bacteriol. 1984 Feb;157(2):363–367. doi: 10.1128/jb.157.2.363-367.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baughman G., Nomura M. Localization of the target site for translational regulation of the L11 operon and direct evidence for translational coupling in Escherichia coli. Cell. 1983 Oct;34(3):979–988. doi: 10.1016/0092-8674(83)90555-x. [DOI] [PubMed] [Google Scholar]
  4. Blumer K. J., Ivey M. R., Steege D. A. Translational control of phage f1 gene expression by differential activities of the gene V, VII, IX and VIII initiation sites. J Mol Biol. 1987 Oct 5;197(3):439–451. doi: 10.1016/0022-2836(87)90557-2. [DOI] [PubMed] [Google Scholar]
  5. Cabezón T., Delcuve G., Faelen M., Desmet L., Bollen A. Polarity of amber mutations in ribosomal protein genes of Escherichia coli. J Bacteriol. 1980 Jan;141(1):41–51. doi: 10.1128/jb.141.1.41-51.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Das A., Yanofsky C. A ribosome binding site sequence is necessary for efficient expression of the distal gene of a translationally-coupled gene pair. Nucleic Acids Res. 1984 Jun 11;12(11):4757–4768. doi: 10.1093/nar/12.11.4757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Freedman L. P., Zengel J. M., Archer R. H., Lindahl L. Autogenous control of the S10 ribosomal protein operon of Escherichia coli: genetic dissection of transcriptional and posttranscriptional regulation. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6516–6520. doi: 10.1073/pnas.84.18.6516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Freedman L. P., Zengel J. M., Lindahl L. Genetic dissection of stringent control and nutritional shift-up response of the Escherichia coli S10 ribosomal protein operon. J Mol Biol. 1985 Oct 20;185(4):701–712. doi: 10.1016/0022-2836(85)90055-5. [DOI] [PubMed] [Google Scholar]
  9. Hansen U., Sharp P. A. Sequences controlling in vitro transcription of SV40 promoters. EMBO J. 1983;2(12):2293–2303. doi: 10.1002/j.1460-2075.1983.tb01737.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lindahl L., Archer R., Zengel J. M. Transcription of the S10 ribosomal protein operon is regulated by an attenuator in the leader. Cell. 1983 May;33(1):241–248. doi: 10.1016/0092-8674(83)90353-7. [DOI] [PubMed] [Google Scholar]
  11. Lindahl L., Zengel J. M. Operon-specific regulation of ribosomal protein synthesis in Escherichia coli. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6542–6546. doi: 10.1073/pnas.76.12.6542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lindahl L., Zengel J. M. Ribosomal genes in Escherichia coli. Annu Rev Genet. 1986;20:297–326. doi: 10.1146/annurev.ge.20.120186.001501. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Mattheakis L. C., Nomura M. Feedback regulation of the spc operon in Escherichia coli: translational coupling and mRNA processing. J Bacteriol. 1988 Oct;170(10):4484–4492. doi: 10.1128/jb.170.10.4484-4492.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  16. Morse D. E., Guertin M. Amber suA mutations which relieve polarity. J Mol Biol. 1972 Feb 14;63(3):605–608. doi: 10.1016/0022-2836(72)90453-6. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Nomura M., Gourse R., Baughman G. Regulation of the synthesis of ribosomes and ribosomal components. Annu Rev Biochem. 1984;53:75–117. doi: 10.1146/annurev.bi.53.070184.000451. [DOI] [PubMed] [Google Scholar]
  19. Oppenheim D. S., Yanofsky C. Translational coupling during expression of the tryptophan operon of Escherichia coli. Genetics. 1980 Aug;95(4):785–795. doi: 10.1093/genetics/95.4.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schmidt B. F., Berkhout B., Overbeek G. P., van Strien A., van Duin J. Determination of the RNA secondary structure that regulates lysis gene expression in bacteriophage MS2. J Mol Biol. 1987 Jun 5;195(3):505–516. doi: 10.1016/0022-2836(87)90179-3. [DOI] [PubMed] [Google Scholar]
  21. Schümperli D., McKenney K., Sobieski D. A., Rosenberg M. Translational coupling at an intercistronic boundary of the Escherichia coli galactose operon. Cell. 1982 Oct;30(3):865–871. doi: 10.1016/0092-8674(82)90291-4. [DOI] [PubMed] [Google Scholar]
  22. Shen P., Zengel J. M., Lindahl L. Secondary structure of the leader transcript from the Escherichia coli S10 ribosomal protein operon. Nucleic Acids Res. 1988 Sep 26;16(18):8905–8924. doi: 10.1093/nar/16.18.8905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sor F., Bolotin-Fukuhara M., Nomura M. Mutational alterations of translational coupling in the L11 ribosomal protein operon of Escherichia coli. J Bacteriol. 1987 Aug;169(8):3495–3507. doi: 10.1128/jb.169.8.3495-3507.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
  25. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  26. Thomas M. S., Bedwell D. M., Nomura M. Regulation of alpha operon gene expression in Escherichia coli. A novel form of translational coupling. J Mol Biol. 1987 Jul 20;196(2):333–345. doi: 10.1016/0022-2836(87)90694-2. [DOI] [PubMed] [Google Scholar]
  27. Yates J. L., Nomura M. E. coli ribosomal protein L4 is a feedback regulatory protein. Cell. 1980 Sep;21(2):517–522. doi: 10.1016/0092-8674(80)90489-4. [DOI] [PubMed] [Google Scholar]
  28. Yates J. L., Nomura M. Feedback regulation of ribosomal protein synthesis in E. coli: localization of the mRNA target sites for repressor action of ribosomal protein L1. Cell. 1981 Apr;24(1):243–249. doi: 10.1016/0092-8674(81)90520-1. [DOI] [PubMed] [Google Scholar]
  29. Zengel J. M., Lindahl L. Transcription of ribosomal genes during a nutritional shift-up of Escherichia coli. J Bacteriol. 1986 Sep;167(3):1095–1097. doi: 10.1128/jb.167.3.1095-1097.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zengel J. M., Mueckl D., Lindahl L. Protein L4 of the E. coli ribosome regulates an eleven gene r protein operon. Cell. 1980 Sep;21(2):523–535. doi: 10.1016/0092-8674(80)90490-0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES