Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Jan;176(2):486–494. doi: 10.1128/jb.176.2.486-494.1994

Expression of both Bacillus subtilis threonyl-tRNA synthetase genes is autogenously regulated.

N Gendron 1, H Putzer 1, M Grunberg-Manago 1
PMCID: PMC205072  PMID: 8288542

Abstract

The "housekeeping" threonyl-tRNA synthetase gene (thrS) of Bacillus subtilis is shown to be transcribed in vivo and in vitro from a single promoter. In vitro, 85% of all messages transcribed from the thrS promoter are terminated at a strong factor-independent terminator localized upstream of the thrS Shine-Dalgarno sequence, within the 305-nucleotide-long leader region. Overexpression of thrS represses transcriptional and translational thrS-lacZ fusions to a similar extent, suggesting that thrS is autoregulated at the transcriptional level. We show that autogenous control does not act at the level of transcription initiation but involves antitermination of the transcription mechanism. thrZ, the second threonyl-tRNA synthetase gene, is also autogenously regulated. However, the ability of the ThrS synthetase to repress thrS as well as thrZ expression is much greater than that of the ThrZ synthetase.

Full text

PDF
486

Images in this article

488Image
on p.488
489Image
on p.489
491Image
on p.491
491Image
on p.491

Selected References

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

  1. Antoniewski C., Savelli B., Stragier P. The spoIIJ gene, which regulates early developmental steps in Bacillus subtilis, belongs to a class of environmentally responsive genes. J Bacteriol. 1990 Jan;172(1):86–93. doi: 10.1128/jb.172.1.86-93.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bardowski J., Ehrlich S. D., Chopin A. Tryptophan biosynthesis genes in Lactococcus lactis subsp. lactis. J Bacteriol. 1992 Oct;174(20):6563–6570. doi: 10.1128/jb.174.20.6563-6570.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernhard K., Schrempf H., Goebel W. Bacteriocin and antibiotic resistance plasmids in Bacillus cereus and Bacillus subtilis. J Bacteriol. 1978 Feb;133(2):897–903. doi: 10.1128/jb.133.2.897-903.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Butler J. S., Springer M., Dondon J., Grunberg-Manago M. Posttranscriptional autoregulation of Escherichia coli threonyl tRNA synthetase expression in vivo. J Bacteriol. 1986 Jan;165(1):198–203. doi: 10.1128/jb.165.1.198-203.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chung C. T., Niemela S. L., Miller R. H. One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2172–2175. doi: 10.1073/pnas.86.7.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Delorme C., Ehrlich S. D., Renault P. Histidine biosynthesis genes in Lactococcus lactis subsp. lactis. J Bacteriol. 1992 Oct;174(20):6571–6579. doi: 10.1128/jb.174.20.6571-6579.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dessen P., Fondrat C., Valencien C., Mugnier C. BISANCE: a French service for access to biomolecular sequence databases. Comput Appl Biosci. 1990 Oct;6(4):355–356. doi: 10.1093/bioinformatics/6.4.355. [DOI] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Grandoni J. A., Zahler S. A., Calvo J. M. Transcriptional regulation of the ilv-leu operon of Bacillus subtilis. J Bacteriol. 1992 May;174(10):3212–3219. doi: 10.1128/jb.174.10.3212-3219.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grundy F. J., Henkin T. M. tRNA as a positive regulator of transcription antitermination in B. subtilis. Cell. 1993 Aug 13;74(3):475–482. doi: 10.1016/0092-8674(93)80049-k. [DOI] [PubMed] [Google Scholar]
  11. Henkin T. M., Glass B. L., Grundy F. J. Analysis of the Bacillus subtilis tyrS gene: conservation of a regulatory sequence in multiple tRNA synthetase genes. J Bacteriol. 1992 Feb;174(4):1299–1306. doi: 10.1128/jb.174.4.1299-1306.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  14. Mechulam Y., Schmitt E., Panvert M., Schmitter J. M., Lapadat-Tapolsky M., Meinnel T., Dessen P., Blanquet S., Fayat G. Methionyl-tRNA synthetase from Bacillus stearothermophilus: structural and functional identities with the Escherichia coli enzyme. Nucleic Acids Res. 1991 Jul 11;19(13):3673–3681. doi: 10.1093/nar/19.13.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ogasawara N., Moriya S., Mazza P. G., Yoshikawa H. Nucleotide sequence and organization of dnaB gene and neighbouring genes on the Bacillus subtilis chromosome. Nucleic Acids Res. 1986 Dec 22;14(24):9989–9999. doi: 10.1093/nar/14.24.9989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Putzer H., Brakhage A. A., Grunberg-Manago M. Independent genes for two threonyl-tRNA synthetases in Bacillus subtilis. J Bacteriol. 1990 Aug;172(8):4593–4602. doi: 10.1128/jb.172.8.4593-4602.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Putzer H., Gendron N., Grunberg-Manago M. Co-ordinate expression of the two threonyl-tRNA synthetase genes in Bacillus subtilis: control by transcriptional antitermination involving a conserved regulatory sequence. EMBO J. 1992 Aug;11(8):3117–3127. doi: 10.1002/j.1460-2075.1992.tb05384.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Seto D. An improved method for sequencing double stranded plasmid DNA from minipreps using DMSO and modified template preparation. Nucleic Acids Res. 1990 Oct 11;18(19):5905–5906. doi: 10.1093/nar/18.19.5905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Springer M., Graffe M., Butler J. S., Grunberg-Manago M. Genetic definition of the translational operator of the threonine-tRNA ligase gene in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4384–4388. doi: 10.1073/pnas.83.12.4384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Springer M., Plumbridge J. A., Butler J. S., Graffe M., Dondon J., Mayaux J. F., Fayat G., Lestienne P., Blanquet S., Grunberg-Manago M. Autogenous control of Escherichia coli threonyl-tRNA synthetase expression in vivo. J Mol Biol. 1985 Sep 5;185(1):93–104. doi: 10.1016/0022-2836(85)90185-8. [DOI] [PubMed] [Google Scholar]
  22. Stragier P., Bonamy C., Karmazyn-Campelli C. Processing of a sporulation sigma factor in Bacillus subtilis: how morphological structure could control gene expression. Cell. 1988 Mar 11;52(5):697–704. doi: 10.1016/0092-8674(88)90407-2. [DOI] [PubMed] [Google Scholar]
  23. Uzan M., Favre R., Brody E. A nuclease that cuts specifically in the ribosome binding site of some T4 mRNAs. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8895–8899. doi: 10.1073/pnas.85.23.8895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Waye M. M., Winter G. A transcription terminator in the 5' non-coding region of the tyrosyl tRNA synthetase gene from Bacillus stearothermophilus. Eur J Biochem. 1986 Aug 1;158(3):505–510. doi: 10.1111/j.1432-1033.1986.tb09783.x. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Zuker M. On finding all suboptimal foldings of an RNA molecule. Science. 1989 Apr 7;244(4900):48–52. doi: 10.1126/science.2468181. [DOI] [PubMed] [Google Scholar]

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

RESOURCES