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. 1989 Sep;171(9):4979–4986. doi: 10.1128/jb.171.9.4979-4986.1989

Structure and expression of the cytochrome aa3 regulatory gene ctaA of Bacillus subtilis.

J P Mueller 1, H W Taber 1
PMCID: PMC210306  PMID: 2549007

Abstract

Mutations that define the ctaA gene of Bacillus subtilis block cytochrome aa3 formation and sporulation. We have recently described the isolation and initial characterization of the ctaA locus. Analysis of in vivo mRNA transcripts by RNase protection experiments located the 5' and 3' termini of the ctaA transcript, confirming a monocistronic structure. By using a nuclease protection assay, an increase in the abundance of steady-state ctaA mRNA was observed during the initiation of sporulation, followed by a decrease during subsequent stages. Transcripts originating from the ctaA gene were most abundant 2.0 h after the end of exponential growth. This pattern of ctaA mRNA accumulation was confirmed by coupling the transcription of the ctaA gene to lacZ in an integrative plasmid vector. Expression of ctaA was not repressed by glucose and was independent of the spoOA and spoOH (sigH) gene products. Postexponential expression was found to be dependent on the product of the strC gene. The expression of ctaA appears to be regulated in a growth stage-specific manner. The transcriptional start site, identified by high-resolution S1 nuclease protection experiments, was preceded by a single sigma A-dependent promoter sequence.

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Selected References

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  1. Albano M., Hahn J., Dubnau D. Expression of competence genes in Bacillus subtilis. J Bacteriol. 1987 Jul;169(7):3110–3117. doi: 10.1128/jb.169.7.3110-3117.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aldea M., Claverie-Martín F., Díaz-Torres M. R., Kushner S. R. Transcript mapping using [35S]DNA probes, trichloroacetate solvent and dideoxy sequencing ladders: a rapid method for identification of transcriptional start points. Gene. 1988 May 15;65(1):101–110. doi: 10.1016/0378-1119(88)90421-0. [DOI] [PubMed] [Google Scholar]
  3. Arrow A. S., Taber H. W. Streptomycin accumulation by Bacillus subtilis requires both a membrane potential and cytochrome aa3. Antimicrob Agents Chemother. 1986 Jan;29(1):141–146. doi: 10.1128/aac.29.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burke J. F. High-sensitivity S1 mapping with single-stranded [32P]DNA probes synthesized from bacteriophage M13mp templates. Gene. 1984 Oct;30(1-3):63–68. doi: 10.1016/0378-1119(84)90105-7. [DOI] [PubMed] [Google Scholar]
  5. CHAIX P., PETIT J. F. Influence du taux de croissance sur la constitution du spectre hématinique de B. subtilis. Biochim Biophys Acta. 1957 Sep;25(3):481–486. doi: 10.1016/0006-3002(57)90517-6. [DOI] [PubMed] [Google Scholar]
  6. Dingman D. W., Rosenkrantz M. S., Sonenshein A. L. Relationship between aconitase gene expression and sporulation in Bacillus subtilis. J Bacteriol. 1987 Jul;169(7):3068–3075. doi: 10.1128/jb.169.7.3068-3075.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doi R. H., Wang L. F. Multiple procaryotic ribonucleic acid polymerase sigma factors. Microbiol Rev. 1986 Sep;50(3):227–243. doi: 10.1128/mr.50.3.227-243.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Errington J. A general method for fusion of the Escherichia coli lacZ gene to chromosomal genes in Bacillus subtilis. J Gen Microbiol. 1986 Nov;132(11):2953–2966. doi: 10.1099/00221287-132-11-2953. [DOI] [PubMed] [Google Scholar]
  9. Farrand S. K., Taber H. W. Physiological effects of menaquinone deficiency in Bacillus subtilis. J Bacteriol. 1973 Sep;115(3):1035–1044. doi: 10.1128/jb.115.3.1035-1044.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feavers I. M., Price V., Moir A. The regulation of the fumarase (citG) gene of Bacillus subtilis 168. Mol Gen Genet. 1988 Mar;211(3):465–471. doi: 10.1007/BF00425702. [DOI] [PubMed] [Google Scholar]
  11. Igo M. M., Losick R. Regulation of a promoter that is utilized by minor forms of RNA polymerase holoenzyme in Bacillus subtilis. J Mol Biol. 1986 Oct 20;191(4):615–624. doi: 10.1016/0022-2836(86)90449-3. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Magnusson K., Philips M. K., Guest J. R., Rutberg L. Nucleotide sequence of the gene for cytochrome b558 of the Bacillus subtilis succinate dehydrogenase complex. J Bacteriol. 1986 Jun;166(3):1067–1071. doi: 10.1128/jb.166.3.1067-1071.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McEnroe A. S., Taber H. W. Correlation between cytochrome aa3 concentrations and streptomycin accumulation in Bacillus subtilis. Antimicrob Agents Chemother. 1984 Oct;26(4):507–512. doi: 10.1128/aac.26.4.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller P., Mueller J., Hill K., Taber H. Transcriptional regulation of a promoter in the men gene cluster of Bacillus subtilis. J Bacteriol. 1988 Jun;170(6):2742–2748. doi: 10.1128/jb.170.6.2742-2748.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  18. Mueller J. P., Taber H. W. Isolation and sequence of ctaA, a gene required for cytochrome aa3 biosynthesis and sporulation in Bacillus subtilis. J Bacteriol. 1989 Sep;171(9):4967–4978. doi: 10.1128/jb.171.9.4967-4978.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Olsson A., Moks T., Uhlén M., Gaal A. B. Uniformly spaced banding pattern in DNA sequencing gels by use of field-strength gradient. J Biochem Biophys Methods. 1984 Nov;10(1-2):83–90. doi: 10.1016/0165-022x(84)90053-8. [DOI] [PubMed] [Google Scholar]
  20. Piggot P. J., Curtis C. A., de Lencastre H. Use of integrational plasmid vectors to demonstrate the polycistronic nature of a transcriptional unit (spoIIA) required for sporulation of Bacillus subtilis. J Gen Microbiol. 1984 Aug;130(8):2123–2136. doi: 10.1099/00221287-130-8-2123. [DOI] [PubMed] [Google Scholar]
  21. Staal S. P., Hoch J. A. Conditional dihydrostreptomycin resistance in Bacillus subtilis. J Bacteriol. 1972 Apr;110(1):202–207. doi: 10.1128/jb.110.1.202-207.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Taber H. Isolation and properties of cytochrome a deficient mutants of Bacillus subtilis. J Gen Microbiol. 1974 Apr;81(2):435–444. doi: 10.1099/00221287-81-2-435. [DOI] [PubMed] [Google Scholar]
  23. Tochikubo K. Changes in terminal respiratory pathways of Bacillus subtilis during germination, outgrowth and vegetative growth. J Bacteriol. 1971 Nov;108(2):652–661. doi: 10.1128/jb.108.2.652-661.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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