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. 1997 Apr;179(8):2587–2594. doi: 10.1128/jb.179.8.2587-2594.1997

Specificity of tRNA-mRNA interactions in Bacillus subtilis tyrS antitermination.

F J Grundy 1, S E Hodil 1, S M Rollins 1, T M Henkin 1
PMCID: PMC179008  PMID: 9098057

Abstract

The Bacillus subtilis tyrS gene, encoding tyrosyl-tRNA synthetase, is a member of the T-box family of genes, which are regulated by control of readthrough of a leader region transcriptional terminator. Readthrough is induced by interaction of the cognate uncharged tRNA with the leader; the system responds to decreased tRNA charging, caused by amino acid limitation or insufficient levels of the aminoacyl-tRNA synthetase. Recognition of the cognate tRNA is mediated by pairing of the anticodon of the tRNA with the specifier sequence of the leader, a codon specifying the appropriate amino acid; a second interaction between the acceptor end of the tRNA and an antiterminator structure is also important. Certain switches of the specifier sequence to a new codon result in a switch in the specificity of the amino acid response, while other switches do not. These effects may reflect additional sequence or structural requirements for the mRNA-tRNA interaction. This study includes investigation of the effects of a large number of specifier sequence switches in tyrS and analysis of structural differences between tRNA(Tyr) and tRNA species which interact inefficiently with the tyrS leader to promote antitermination.

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Selected References

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  1. Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Garrity D. B., Zahler S. A. Mutations in the gene for a tRNA that functions as a regulator of a transcriptional attenuator in Bacillus subtilis. Genetics. 1994 Jul;137(3):627–636. doi: 10.1093/genetics/137.3.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Garrity D. B., Zahler S. A. The Bacillus subtilis ochre suppressor sup-3 is located in an operon of seven tRNA genes. J Bacteriol. 1993 Oct;175(20):6512–6517. doi: 10.1128/jb.175.20.6512-6517.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Grandoni J. A., Fulmer S. B., Brizzio V., Zahler S. A., Calvo J. M. Regions of the Bacillus subtilis ilv-leu operon involved in regulation by leucine. J Bacteriol. 1993 Dec;175(23):7581–7593. doi: 10.1128/jb.175.23.7581-7593.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Green C. J., Vold B. S. A cluster of nine tRNA genes between ribosomal gene operons in Bacillus subtilis. J Bacteriol. 1992 May;174(10):3147–3151. doi: 10.1128/jb.174.10.3147-3151.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Green C. J., Vold B. S. Sequence analysis of a cluster of twenty-one tRNA genes in Bacillus subtilis. Nucleic Acids Res. 1983 Aug 25;11(16):5763–5774. doi: 10.1093/nar/11.16.5763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Grundy F. J., Henkin T. M. Conservation of a transcription antitermination mechanism in aminoacyl-tRNA synthetase and amino acid biosynthesis genes in gram-positive bacteria. J Mol Biol. 1994 Jan 14;235(2):798–804. doi: 10.1006/jmbi.1994.1038. [DOI] [PubMed] [Google Scholar]
  8. Grundy F. J., Henkin T. M. The rpsD gene, encoding ribosomal protein S4, is autogenously regulated in Bacillus subtilis. J Bacteriol. 1991 Aug;173(15):4595–4602. doi: 10.1128/jb.173.15.4595-4602.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Grundy F. J., Rollins S. M., Henkin T. M. Interaction between the acceptor end of tRNA and the T box stimulates antitermination in the Bacillus subtilis tyrS gene: a new role for the discriminator base. J Bacteriol. 1994 Aug;176(15):4518–4526. doi: 10.1128/jb.176.15.4518-4526.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grundy F. J., Waters D. A., Allen S. H., Henkin T. M. Regulation of the Bacillus subtilis acetate kinase gene by CcpA. J Bacteriol. 1993 Nov;175(22):7348–7355. doi: 10.1128/jb.175.22.7348-7355.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hasegawa T., Miyano M., Himeno H., Sano Y., Kimura K., Shimizu M. Identity determinants of E. coli threonine tRNA. Biochem Biophys Res Commun. 1992 Apr 15;184(1):478–484. doi: 10.1016/0006-291x(92)91219-g. [DOI] [PubMed] [Google Scholar]
  13. Henkin T. M., Chambliss G. H. Genetic mapping of a mutation causing an alteration in Bacillus subtilis ribosomal protein S4. Mol Gen Genet. 1984;193(2):364–369. doi: 10.1007/BF00330694. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Henkin T. M. tRNA-directed transcription antitermination. Mol Microbiol. 1994 Aug;13(3):381–387. doi: 10.1111/j.1365-2958.1994.tb00432.x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Ladner J. E., Jack A., Robertus J. D., Brown R. S., Rhodes D., Clark B. F., Klug A. Structure of yeast phenylalanine transfer RNA at 2.5 A resolution. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4414–4418. doi: 10.1073/pnas.72.11.4414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marta P. T., Ladner R. D., Grandoni J. A. A CUC triplet confers leucine-dependent regulation of the Bacillus subtilis ilv-leu operon. J Bacteriol. 1996 Apr;178(7):2150–2153. doi: 10.1128/jb.178.7.2150-2153.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Nakano M. M., Zuber P. Cloning and characterization of srfB, a regulatory gene involved in surfactin production and competence in Bacillus subtilis. J Bacteriol. 1989 Oct;171(10):5347–5353. doi: 10.1128/jb.171.10.5347-5353.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ogasawara N., Moriya S., Yoshikawa H. Structure and organization of rRNA operons in the region of the replication origin of the Bacillus subtilis chromosome. Nucleic Acids Res. 1983 Sep 24;11(18):6301–6318. doi: 10.1093/nar/11.18.6301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Perret V., Garcia A., Grosjean H., Ebel J. P., Florentz C., Giegé R. Relaxation of a transfer RNA specificity by removal of modified nucleotides. Nature. 1990 Apr 19;344(6268):787–789. doi: 10.1038/344787a0. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Putzer H., Laalami S., Brakhage A. A., Condon C., Grunberg-Manago M. Aminoacyl-tRNA synthetase gene regulation in Bacillus subtilis: induction, repression and growth-rate regulation. Mol Microbiol. 1995 May;16(4):709–718. doi: 10.1111/j.1365-2958.1995.tb02432.x. [DOI] [PubMed] [Google Scholar]
  25. Steinmetz M., Richter R. Plasmids designed to alter the antibiotic resistance expressed by insertion mutations in Bacillus subtilis, through in vivo recombination. Gene. 1994 May 3;142(1):79–83. doi: 10.1016/0378-1119(94)90358-1. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Wawrousek E. F., Hansen J. N. Structure and organization of a cluster of sic tRNA genes in the space between tandem ribosomal RNA gene sets in Bacillus subtilis. J Biol Chem. 1983 Jan 10;258(1):291–298. [PubMed] [Google Scholar]
  28. Wawrousek E. F., Narasimhan N., Hansen J. N. Two large clusters with thirty-seven transfer RNA genes adjacent to ribosomal RNA gene sets in Bacillus subtilis. Sequence and organization of trrnD and trrnE gene clusters. J Biol Chem. 1984 Mar 25;259(6):3694–3702. [PubMed] [Google Scholar]
  29. Zuber P., Losick R. Role of AbrB in Spo0A- and Spo0B-dependent utilization of a sporulation promoter in Bacillus subtilis. J Bacteriol. 1987 May;169(5):2223–2230. doi: 10.1128/jb.169.5.2223-2230.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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