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. 1994 Apr;176(8):2427–2434. doi: 10.1128/jb.176.8.2427-2434.1994

Characterization of the sigD transcription unit of Bacillus subtilis.

L M Márquez-Magaña 1, M J Chamberlin 1
PMCID: PMC205368  PMID: 8157612

Abstract

The sigma D factor of Bacillus subtilis is required for the transcription of the flagellin and motility genes as well as for wild-type chemotaxis. Southern blot and sequence analyses demonstrate that the structural gene for sigma D, sigD, is located immediately downstream of a region of DNA originally identified as the chemotaxis (che) locus and now renamed the fla/che region. In fact, sigD appears to be part of a very large operon (> 26 kb) containing genes which encode structural proteins that form the hook-basal body complex as well as regulatory proteins required for chemotaxis. Transposon insertions up to 24 kb upstream of sigD, within several of the genes for the hook-basal body components, give rise to only a moderate decrease in sigD expression. The transposon insertions, however, block sigma D activity as demonstrated by the lack of flagellin expression in strains bearing these insertions. These effects appear to arise from two types of regulation. In cis the transposon insertions appear to introduce a partial block to transcription of sigD from upstream promoter elements; in trans they disrupt genes whose gene products are required for sigma D activity. It appears that sigD transcription is initiated, at least in part, by a promoter many kilobases upstream of its translation start site and that transcription of the flagellin gene by sigma D is dependent on the formation of a functional hook-basal body complex. The possibility that sigD is part of the fla/che operon was further tested by the integration of an insertion plasmid, containing strong transcription terminators, 1.6 and 24 kb upstream of the sigD gene. In both cases, the introduction of the terminators resulted in a greater decrease of sigD expression than was caused by the plasmid sequences alone. These results indicate that wild-type transcription of sigD is dependent on promoter sequences > 24kb upstream of its structural gene and that the entire fla/che region forms a single operon.

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Selected References

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  1. Albertini A. M., Caramori T., Crabb W. D., Scoffone F., Galizzi A. The flaA locus of Bacillus subtilis is part of a large operon coding for flagellar structures, motility functions, and an ATPase-like polypeptide. J Bacteriol. 1991 Jun;173(11):3573–3579. doi: 10.1128/jb.173.11.3573-3579.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Arnosti D. N., Chamberlin M. J. Secondary sigma factor controls transcription of flagellar and chemotaxis genes in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Feb;86(3):830–834. doi: 10.1073/pnas.86.3.830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bartlett D. H., Frantz B. B., Matsumura P. Flagellar transcriptional activators FlbB and FlaI: gene sequences and 5' consensus sequences of operons under FlbB and FlaI control. J Bacteriol. 1988 Apr;170(4):1575–1581. doi: 10.1128/jb.170.4.1575-1581.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bischoff D. S., Ordal G. W. Sequence and characterization of Bacillus subtilis CheB, a homolog of Escherichia coli CheY, and its role in a different mechanism of chemotaxis. J Biol Chem. 1991 Jul 5;266(19):12301–12305. [PubMed] [Google Scholar]
  6. Brosius J., Dull T. J., Sleeter D. D., Noller H. F. Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. J Mol Biol. 1981 May 15;148(2):107–127. doi: 10.1016/0022-2836(81)90508-8. [DOI] [PubMed] [Google Scholar]
  7. Brosius J. Toxicity of an overproduced foreign gene product in Escherichia coli and its use in plasmid vectors for the selection of transcription terminators. Gene. 1984 Feb;27(2):161–172. doi: 10.1016/0378-1119(84)90137-9. [DOI] [PubMed] [Google Scholar]
  8. Carlomagno M. S., Chiariotti L., Alifano P., Nappo A. G., Bruni C. B. Structure and function of the Salmonella typhimurium and Escherichia coli K-12 histidine operons. J Mol Biol. 1988 Oct 5;203(3):585–606. doi: 10.1016/0022-2836(88)90194-5. [DOI] [PubMed] [Google Scholar]
  9. Ferrari E., Henner D. J., Hoch J. A. Isolation of Bacillus subtilis genes from a charon 4A library. J Bacteriol. 1981 Apr;146(1):430–432. doi: 10.1128/jb.146.1.430-432.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fuhrer D. K., Ordal G. W. Bacillus subtilis CheN, a homolog of CheA, the central regulator of chemotaxis in Escherichia coli. J Bacteriol. 1991 Dec;173(23):7443–7448. doi: 10.1128/jb.173.23.7443-7448.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gillen K. L., Hughes K. T. Molecular characterization of flgM, a gene encoding a negative regulator of flagellin synthesis in Salmonella typhimurium. J Bacteriol. 1991 Oct;173(20):6453–6459. doi: 10.1128/jb.173.20.6453-6459.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gillen K. L., Hughes K. T. Negative regulatory loci coupling flagellin synthesis to flagellar assembly in Salmonella typhimurium. J Bacteriol. 1991 Apr;173(7):2301–2310. doi: 10.1128/jb.173.7.2301-2310.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gilman M. Z., Chamberlin M. J. Developmental and genetic regulation of Bacillus subtilis genes transcribed by sigma 28-RNA polymerase. Cell. 1983 Nov;35(1):285–293. doi: 10.1016/0092-8674(83)90231-3. [DOI] [PubMed] [Google Scholar]
  14. Hanlon D. W., Márquez-Magaña L. M., Carpenter P. B., Chamberlin M. J., Ordal G. W. Sequence and characterization of Bacillus subtilis CheW. J Biol Chem. 1992 Jun 15;267(17):12055–12060. [PubMed] [Google Scholar]
  15. Helmann J. D. Alternative sigma factors and the regulation of flagellar gene expression. Mol Microbiol. 1991 Dec;5(12):2875–2882. doi: 10.1111/j.1365-2958.1991.tb01847.x. [DOI] [PubMed] [Google Scholar]
  16. Helmann J. D., Chamberlin M. J. Structure and function of bacterial sigma factors. Annu Rev Biochem. 1988;57:839–872. doi: 10.1146/annurev.bi.57.070188.004203. [DOI] [PubMed] [Google Scholar]
  17. Helmann J. D., Masiarz F. R., Chamberlin M. J. Isolation and characterization of the Bacillus subtilis sigma 28 factor. J Bacteriol. 1988 Apr;170(4):1560–1567. doi: 10.1128/jb.170.4.1560-1567.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Helmann J. D., Márquez L. M., Chamberlin M. J. Cloning, sequencing, and disruption of the Bacillus subtilis sigma 28 gene. J Bacteriol. 1988 Apr;170(4):1568–1574. doi: 10.1128/jb.170.4.1568-1574.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jones C. J., Macnab R. M. Flagellar assembly in Salmonella typhimurium: analysis with temperature-sensitive mutants. J Bacteriol. 1990 Mar;172(3):1327–1339. doi: 10.1128/jb.172.3.1327-1339.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Komeda Y. Transcriptional control of flagellar genes in Escherichia coli K-12. J Bacteriol. 1986 Dec;168(3):1315–1318. doi: 10.1128/jb.168.3.1315-1318.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kutsukake K., Ohya Y., Iino T. Transcriptional analysis of the flagellar regulon of Salmonella typhimurium. J Bacteriol. 1990 Feb;172(2):741–747. doi: 10.1128/jb.172.2.741-747.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Losick R., Youngman P., Piggot P. J. Genetics of endospore formation in Bacillus subtilis. Annu Rev Genet. 1986;20:625–669. doi: 10.1146/annurev.ge.20.120186.003205. [DOI] [PubMed] [Google Scholar]
  23. Massie H. R., Zimm B. H. Molecular weight of the DNA in the chromosomes of E. coli and B. subtilis. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1636–1641. doi: 10.1073/pnas.54.6.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mirel D. B., Chamberlin M. J. The Bacillus subtilis flagellin gene (hag) is transcribed by the sigma 28 form of RNA polymerase. J Bacteriol. 1989 Jun;171(6):3095–3101. doi: 10.1128/jb.171.6.3095-3101.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mirel D. B., Lustre V. M., Chamberlin M. J. An operon of Bacillus subtilis motility genes transcribed by the sigma D form of RNA polymerase. J Bacteriol. 1992 Jul;174(13):4197–4204. doi: 10.1128/jb.174.13.4197-4204.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Márquez L. M., Helmann J. D., Ferrari E., Parker H. M., Ordal G. W., Chamberlin M. J. Studies of sigma D-dependent functions in Bacillus subtilis. J Bacteriol. 1990 Jun;172(6):3435–3443. doi: 10.1128/jb.172.6.3435-3443.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ohnishi K., Kutsukake K., Suzuki H., Iino T. Gene fliA encodes an alternative sigma factor specific for flagellar operons in Salmonella typhimurium. Mol Gen Genet. 1990 Apr;221(2):139–147. doi: 10.1007/BF00261713. [DOI] [PubMed] [Google Scholar]
  28. Ordal G. W., Parker H. M., Kirby J. R. Complementation and characterization of chemotaxis mutants of Bacillus subtilis. J Bacteriol. 1985 Nov;164(2):802–810. doi: 10.1128/jb.164.2.802-810.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schmid M. B., Roth J. R. Internal promoters of the his operon in Salmonella typhimurium. J Bacteriol. 1983 Feb;153(2):1114–1119. doi: 10.1128/jb.153.2.1114-1119.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Suzuki T., Komeda Y. Incomplete flagellar structures in Escherichia coli mutants. J Bacteriol. 1981 Feb;145(2):1036–1041. doi: 10.1128/jb.145.2.1036-1041.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zuberi A. R., Bischoff D. S., Ordal G. W. Nucleotide sequence and characterization of a Bacillus subtilis gene encoding a flagellar switch protein. J Bacteriol. 1991 Jan;173(2):710–719. doi: 10.1128/jb.173.2.710-719.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zuberi A. R., Ying C. W., Parker H. M., Ordal G. W. Transposon Tn917lacZ mutagenesis of Bacillus subtilis: identification of two new loci required for motility and chemotaxis. J Bacteriol. 1990 Dec;172(12):6841–6848. doi: 10.1128/jb.172.12.6841-6848.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zuberi A. R., Ying C. W., Weinreich M. R., Ordal G. W. Transcriptional organization of a cloned chemotaxis locus of Bacillus subtilis. J Bacteriol. 1990 Apr;172(4):1870–1876. doi: 10.1128/jb.172.4.1870-1876.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zuberi A. R., Ying C., Bischoff D. S., Ordal G. W. Gene-protein relationships in the flagellar hook-basal body complex of Bacillus subtilis: sequences of the flgB, flgC, flgG, fliE and fliF genes. Gene. 1991 May 15;101(1):23–31. doi: 10.1016/0378-1119(91)90220-6. [DOI] [PubMed] [Google Scholar]

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