Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1992 Aug;11(8):3117–3127. doi: 10.1002/j.1460-2075.1992.tb05384.x

Co-ordinate expression of the two threonyl-tRNA synthetase genes in Bacillus subtilis: control by transcriptional antitermination involving a conserved regulatory sequence.

H Putzer 1, N Gendron 1, M Grunberg-Manago 1
PMCID: PMC556796  PMID: 1379177

Abstract

In Bacillus subtilis, two genes, thrS and thrZ, encode distinct threonyl-tRNA synthetase enzymes. Normally, only the thrS gene is expressed. Here we show that either gene, thrS or thrZ, is sufficient for normal cell growth and sporulation. Reducing the intracellular ThrS protein concentration induces thrZ expression in a dose-compensatory manner. Starvation for threonine simultaneously induces thrZ and stimulates thrS expression. The 5'-leader sequences of thrS and thrZ contain, respectively, one and three transcription terminators preceded by a conserved sequence. We show that this sequence is essential for the regulation of thrS via a transcriptional antitermination mechanism. We propose that both genes, thrS and thrZ, are regulated by the same mechanism such that the additional regulatory domains present before thrZ account for its non-expression. In contrast to Escherichia coli, structurally similar regulatory domains, i.e. the consensus sequence preceding a terminator structure, are found in the leader regions of most aminoacyl-tRNA synthetase genes of Gram-positive bacteria. This suggests that they are regulated by a common mechanism.

Full text

PDF
3117

Images in this article

Selected References

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

  1. Akins R. A., Lambowitz A. M. A protein required for splicing group I introns in Neurospora mitochondria is mitochondrial tyrosyl-tRNA synthetase or a derivative thereof. Cell. 1987 Jul 31;50(3):331–345. doi: 10.1016/0092-8674(87)90488-0. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Barstow D. A., Sharman A. F., Atkinson T., Minton N. P. Cloning and complete nucleotide sequence of the Bacillus stearothermophilus tryptophanyl tRNA synthetase gene. Gene. 1986;46(1):37–45. doi: 10.1016/0378-1119(86)90164-2. [DOI] [PubMed] [Google Scholar]
  4. Brakhage A. A., Wozny M., Putzer H. Structure and nucleotide sequence of the Bacillus subtilis phenylalanyl-tRNA synthetase genes. Biochimie. 1990 Oct;72(10):725–734. doi: 10.1016/0300-9084(90)90157-c. [DOI] [PubMed] [Google Scholar]
  5. Brevet A., Chen J., Lévêque F., Plateau P., Blanquet S. In vivo synthesis of adenylylated bis(5'-nucleosidyl) tetraphosphates (Ap4N) by Escherichia coli aminoacyl-tRNA synthetases. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8275–8279. doi: 10.1073/pnas.86.21.8275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chambers S. P., Prior S. E., Barstow D. A., Minton N. P. The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene. 1988 Aug 15;68(1):139–149. doi: 10.1016/0378-1119(88)90606-3. [DOI] [PubMed] [Google Scholar]
  7. Chow K. C., Wong J. T. Cloning and nucleotide sequence of the structural gene coding for Bacillus subtilis tryptophanyl-tRNA synthetase. Gene. 1988 Dec 20;73(2):537–543. doi: 10.1016/0378-1119(88)90518-5. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Crutz A. M., Steinmetz M., Aymerich S., Richter R., Le Coq D. Induction of levansucrase in Bacillus subtilis: an antitermination mechanism negatively controlled by the phosphotransferase system. J Bacteriol. 1990 Feb;172(2):1043–1050. doi: 10.1128/jb.172.2.1043-1050.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Debarbouille M., Arnaud M., Fouet A., Klier A., Rapoport G. The sacT gene regulating the sacPA operon in Bacillus subtilis shares strong homology with transcriptional antiterminators. J Bacteriol. 1990 Jul;172(7):3966–3973. doi: 10.1128/jb.172.7.3966-3973.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Ferrara P., Duchange N., Zakin M. M., Cohen G. N. Internal homologies in the two aspartokinase-homoserine dehydrogenases of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1984 May;81(10):3019–3023. doi: 10.1073/pnas.81.10.3019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Glaser P., Kunst F., Débarbouillé M., Vertès A., Danchin A., Dedonder R. A gene encoding a tyrosine tRNA synthetase is located near sacS in Bacillus subtilis. DNA Seq. 1991;1(4):251–261. doi: 10.3109/10425179109020780. [DOI] [PubMed] [Google Scholar]
  15. Gollnick P., Ishino S., Kuroda M. I., Henner D. J., Yanofsky C. The mtr locus is a two-gene operon required for transcription attenuation in the trp operon of Bacillus subtilis. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8726–8730. doi: 10.1073/pnas.87.22.8726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Haworth S. R., Brown L. R. Genetic analysis of ribonucleic acid polymerase mutants of Bacillus subtilis. J Bacteriol. 1973 Apr;114(1):103–113. doi: 10.1128/jb.114.1.103-113.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Herbert C. J., Labouesse M., Dujardin G., Slonimski P. P. The NAM2 proteins from S. cerevisiae and S. douglasii are mitochondrial leucyl-tRNA synthetases, and are involved in mRNA splicing. EMBO J. 1988 Feb;7(2):473–483. doi: 10.1002/j.1460-2075.1988.tb02835.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hirshfield I. N., Bloch P. L., Van Bogelen R. A., Neidhardt F. C. Multiple forms of lysyl-transfer ribonucleic acid synthetase in Escherichia coli. J Bacteriol. 1981 Apr;146(1):345–351. doi: 10.1128/jb.146.1.345-351.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hirshfield I. N., Tenreiro R., Vanbogelen R. A., Neidhardt F. C. Escherichia coli K-12 lysyl-tRNA synthetase mutant with a novel reversion pattern. J Bacteriol. 1984 May;158(2):615–620. doi: 10.1128/jb.158.2.615-620.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Horinouchi S., Weisblum B. Posttranscriptional modification of mRNA conformation: mechanism that regulates erythromycin-induced resistance. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7079–7083. doi: 10.1073/pnas.77.12.7079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Houman F., Diaz-Torres M. R., Wright A. Transcriptional antitermination in the bgl operon of E. coli is modulated by a specific RNA binding protein. Cell. 1990 Sep 21;62(6):1153–1163. doi: 10.1016/0092-8674(90)90392-r. [DOI] [PubMed] [Google Scholar]
  24. Iuchi S., Cole S. T., Lin E. C. Multiple regulatory elements for the glpA operon encoding anaerobic glycerol-3-phosphate dehydrogenase and the glpD operon encoding aerobic glycerol-3-phosphate dehydrogenase in Escherichia coli: further characterization of respiratory control. J Bacteriol. 1990 Jan;172(1):179–184. doi: 10.1128/jb.172.1.179-184.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jaskunas S. R., Lindahl L., Nomura M. Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature. 1975 Oct 9;257(5526):458–462. doi: 10.1038/257458a0. [DOI] [PubMed] [Google Scholar]
  26. Kieny M. P., Lathe R., Lecocq J. P. New versatile cloning and sequencing vectors based on bacteriophage M13. Gene. 1983 Dec;26(1):91–99. doi: 10.1016/0378-1119(83)90039-2. [DOI] [PubMed] [Google Scholar]
  27. Kreft J., Bernhard K., Goebel W. Recombinant plasmids capable to replication in B. subtilis and E. coli. Mol Gen Genet. 1978 Jun 1;162(1):59–67. doi: 10.1007/BF00333851. [DOI] [PubMed] [Google Scholar]
  28. Kuritzkes D. R., Zhang X. Y., Lin E. C. Use of phi(glp-lac) in studies of respiratory regulation of the Escherichia coli anaerobic sn-glycerol-3-phosphate dehydrogenase genes (glpAB). J Bacteriol. 1984 Feb;157(2):591–598. doi: 10.1128/jb.157.2.591-598.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kuroda M. I., Henner D., Yanofsky C. cis-acting sites in the transcript of the Bacillus subtilis trp operon regulate expression of the operon. J Bacteriol. 1988 Jul;170(7):3080–3088. doi: 10.1128/jb.170.7.3080-3088.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Lévêque F., Gazeau M., Fromant M., Blanquet S., Plateau P. Control of Escherichia coli lysyl-tRNA synthetase expression by anaerobiosis. J Bacteriol. 1991 Dec;173(24):7903–7910. doi: 10.1128/jb.173.24.7903-7910.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lévêque F., Plateau P., Dessen P., Blanquet S. Homology of lysS and lysU, the two Escherichia coli genes encoding distinct lysyl-tRNA synthetase species. Nucleic Acids Res. 1990 Jan 25;18(2):305–312. doi: 10.1093/nar/18.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. 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]
  35. Parsot C., Cohen G. N. Cloning and nucleotide sequence of the Bacillus subtilis hom gene coding for homoserine dehydrogenase. Structural and evolutionary relationships with Escherichia coli aspartokinases-homoserine dehydrogenases I and II. J Biol Chem. 1988 Oct 15;263(29):14654–14660. [PubMed] [Google Scholar]
  36. Perego M., Spiegelman G. B., Hoch J. A. Structure of the gene for the transition state regulator, abrB: regulator synthesis is controlled by the spo0A sporulation gene in Bacillus subtilis. Mol Microbiol. 1988 Nov;2(6):689–699. doi: 10.1111/j.1365-2958.1988.tb00079.x. [DOI] [PubMed] [Google Scholar]
  37. Price C. W., Doi R. H. Genetic mapping of rpoD implicates the major sigma factor of Bacillus subtilis RNA polymerase in sporulation initiation. Mol Gen Genet. 1985;201(1):88–95. doi: 10.1007/BF00397991. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. 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]
  40. Schaeffer P., Millet J., Aubert J. P. Catabolic repression of bacterial sporulation. Proc Natl Acad Sci U S A. 1965 Sep;54(3):704–711. doi: 10.1073/pnas.54.3.704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Van Vliet F., Cunin R., Jacobs A., Piette J., Gigot D., Lauwereys M., Piérard A., Glansdorff N. Evolutionary divergence of genes for ornithine and aspartate carbamoyl-transferases--complete sequence and mode of regulation of the Escherichia coli argF gene; comparison of argF with argI and pyrB. Nucleic Acids Res. 1984 Aug 10;12(15):6277–6289. doi: 10.1093/nar/12.15.6277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. 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 The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES