Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Feb;169(2):461–469. doi: 10.1128/jb.169.2.461-469.1987

Effects of plasmid propagation of a sporulation promoter on promoter utilization and sporulation in Bacillus subtilis.

P Zuber, J M Healy, R Losick
PMCID: PMC211799  PMID: 3027029

Abstract

Transcription of the sporulation gene spoVG of Bacillus subtilis is induced at the onset of spore formation and depends on the products of the regulatory genes spoOA, spoOB, and spoOH. We describe two effects of propagating the promoter region of spoVG on a multicopy plasmid replicon in B. subtilis cells. One effect is that transcription from the plasmid-borne spoVG promoter is altered with respect to the time of its induction and the dependence on spoO gene products. An example of this effect is that plasmid propagation was observed to relieve substantially the inhibitory effect of a mutation in spoOH, the spoO gene upon which spoVG promoter activity is most strongly dependent. We present results which suggest that propagation on a plasmid replicon causes an alteration in the conformation of spoVG promoter DNA which somehow compensates for the defective spoOH gene product. Plasmid propagation did not, however, entirely eliminate the requirement for the spoOH gene product; little or no spoVG-directed RNA synthesis was observed in cells bearing a putative spoOH deletion mutation, a finding which indicates that SpoOH protein plays an indispensable role in spoVG promoter utilization. Another effect of propagating the promoter region of spoVG on a multicopy plasmid is to inhibit sporulation. S1 nuclease mapping experiments suggest that amplification of spoVG on a multicopy plasmid causes the titration of a transcription factor or minor form of RNA polymerase holoenzyme required for utilization of one of the two overlapping promoters which comprise the spoVG transcription initiation region.

Full text

PDF
461

Images in this article

Selected References

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

  1. Banner C. D., Moran C. P., Jr, Losick R. Deletion analysis of a complex promoter for a developmentally regulated gene from Bacillus subtilis. J Mol Biol. 1983 Aug 5;168(2):351–365. doi: 10.1016/s0022-2836(83)80023-0. [DOI] [PubMed] [Google Scholar]
  2. Binnie C., Lampe M., Losick R. Gene encoding the sigma 37 species of RNA polymerase sigma factor from Bacillus subtilis. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5943–5947. doi: 10.1073/pnas.83.16.5943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouvier J., Stragier P., Bonamy C., Szulmajster J. Nucleotide sequence of the spo0B gene of Bacillus subtilis and regulation of its expression. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7012–7016. doi: 10.1073/pnas.81.22.7012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ferrari F. A., Trach K., Hoch J. A. Sequence analysis of the spo0B locus reveals a polycistronic transcription unit. J Bacteriol. 1985 Feb;161(2):556–562. doi: 10.1128/jb.161.2.556-562.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ferrari F. A., Trach K., LeCoq D., Spence J., Ferrari E., Hoch J. A. Characterization of the spo0A locus and its deduced product. Proc Natl Acad Sci U S A. 1985 May;82(9):2647–2651. doi: 10.1073/pnas.82.9.2647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gryczan T. J., Dubnau D. Construction and properties of chimeric plasmids in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1428–1432. doi: 10.1073/pnas.75.3.1428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gryczan T., Shivakumar A. G., Dubnau D. Characterization of chimeric plasmid cloning vehicles in Bacillus subtilis. J Bacteriol. 1980 Jan;141(1):246–253. doi: 10.1128/jb.141.1.246-253.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haldenwang W. G., Losick R. A modified RNA polymerase transcribes a cloned gene under sporulation control in Bacillus subtilis. Nature. 1979 Nov 15;282(5736):256–260. doi: 10.1038/282256a0. [DOI] [PubMed] [Google Scholar]
  9. Hoch J. A. Genetics of bacterial sporulation. Adv Genet. 1976;18:69–98. doi: 10.1016/s0065-2660(08)60437-x. [DOI] [PubMed] [Google Scholar]
  10. Johnson W. C., Moran C. P., Jr, Losick R. Two RNA polymerase sigma factors from Bacillus subtilis discriminate between overlapping promoters for a developmentally regulated gene. Nature. 1983 Apr 28;302(5911):800–804. doi: 10.1038/302800a0. [DOI] [PubMed] [Google Scholar]
  11. Losick R., Pero J. Cascades of Sigma factors. Cell. 1981 Sep;25(3):582–584. doi: 10.1016/0092-8674(81)90164-1. [DOI] [PubMed] [Google Scholar]
  12. Malan T. P., Kolb A., Buc H., McClure W. R. Mechanism of CRP-cAMP activation of lac operon transcription initiation activation of the P1 promoter. J Mol Biol. 1984 Dec 25;180(4):881–909. doi: 10.1016/0022-2836(84)90262-6. [DOI] [PubMed] [Google Scholar]
  13. McClure W. R. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem. 1985;54:171–204. doi: 10.1146/annurev.bi.54.070185.001131. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Menzel R., Gellert M. Regulation of the genes for E. coli DNA gyrase: homeostatic control of DNA supercoiling. Cell. 1983 Aug;34(1):105–113. doi: 10.1016/0092-8674(83)90140-x. [DOI] [PubMed] [Google Scholar]
  16. Mitani T., Heinze J. E., Freese E. Induction of sporulation in Bacillus subtilis by decoyinine or hadacidin. Biochem Biophys Res Commun. 1977 Aug 8;77(3):1118–1125. doi: 10.1016/s0006-291x(77)80094-6. [DOI] [PubMed] [Google Scholar]
  17. Moran C. P., Jr, Lang N., Banner C. D., Haldenwang W. G., Losick R. Promoter for a developmentally regulated gene in Bacillus subtilis. Cell. 1981 Sep;25(3):783–791. doi: 10.1016/0092-8674(81)90186-0. [DOI] [PubMed] [Google Scholar]
  18. Ollington J. F., Haldenwang W. G., Huynh T. V., Losick R. Developmentally regulated transcription in a cloned segment of the Bacillus subtilis chromosome. J Bacteriol. 1981 Aug;147(2):432–442. doi: 10.1128/jb.147.2.432-442.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Penn M. D., Thireos G., Greer H. Temporal analysis of general control of amino acid biosynthesis in Saccharomyces cerevisiae: role of positive regulatory genes in initiation and maintenance of mRNA derepression. Mol Cell Biol. 1984 Mar;4(3):520–528. doi: 10.1128/mcb.4.3.520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Piggot P. J., Coote J. G. Genetic aspects of bacterial endospore formation. Bacteriol Rev. 1976 Dec;40(4):908–962. doi: 10.1128/br.40.4.908-962.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ramakrishna N., Dubnau E., Smith I. The complete DNA sequence and regulatory regions of the Bacillus licheniformis spoOH gene. Nucleic Acids Res. 1984 Feb 24;12(4):1779–1790. doi: 10.1093/nar/12.4.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rosenkrantz M. S., Dingman D. W., Sonenshein A. L. Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine. J Bacteriol. 1985 Oct;164(1):155–164. doi: 10.1128/jb.164.1.155-164.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sanzey B. Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase. J Bacteriol. 1979 Apr;138(1):40–47. doi: 10.1128/jb.138.1.40-47.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shivakumar A. G., Dubnau D. Differential effect of hydroxyurea on the replication of plasmid and chromosomal DNA in Bacillus subtilis. J Bacteriol. 1978 Dec;136(3):1205–1207. doi: 10.1128/jb.136.3.1205-1207.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith G. E., Summers M. D. The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980 Nov 15;109(1):123–129. doi: 10.1016/0003-2697(80)90019-6. [DOI] [PubMed] [Google Scholar]
  26. Smith G. R. DNA supercoiling: another level for regulating gene expression. Cell. 1981 Jun;24(3):599–600. doi: 10.1016/0092-8674(81)90085-4. [DOI] [PubMed] [Google Scholar]
  27. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  28. Trach K. A., Chapman J. W., Piggot P. J., Hoch J. A. Deduced product of the stage 0 sporulation gene spo0F shares homology with the Spo0A, OmpR, and SfrA proteins. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7260–7264. doi: 10.1073/pnas.82.21.7260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weir J., Dubnau E., Ramakrishna N., Smith I. Bacillus subtilis spo0H gene. J Bacteriol. 1984 Feb;157(2):405–412. doi: 10.1128/jb.157.2.405-412.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Young M. Gene amplification in Bacillus subtilis. J Gen Microbiol. 1984 Jul;130(7):1613–1621. doi: 10.1099/00221287-130-7-1613. [DOI] [PubMed] [Google Scholar]
  31. Zuber P., Losick R. Use of a lacZ fusion to study the role of the spoO genes of Bacillus subtilis in developmental regulation. Cell. 1983 Nov;35(1):275–283. doi: 10.1016/0092-8674(83)90230-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES