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. 1981 Aug;147(2):432–442. doi: 10.1128/jb.147.2.432-442.1981

Developmentally regulated transcription in a cloned segment of the Bacillus subtilis chromosome.

J F Ollington, W G Haldenwang, T V Huynh, R Losick
PMCID: PMC216062  PMID: 6790515

Abstract

We describe a model system for studying developmentally regulated transcription during spore formation in Bacillus subtilis. This model system is a cloned cluster of genes known as 0.4 kb, ctc, and veg from the purA-cysA region of the B. subtilis chromosome. Each gene exhibited a distinct pattern of transcription in cells growing in glucose medium and in cells deprived of nutrients in sporulation medium. The 0.4 kb gene was transcribed at a low level in growing cells but was actively transcribed during nutrient deprivation in sporulation medium. This ribonucleic acid (RNA) synthesis was dependent upon the products of five B. subtilis genes that are involved in the initiation of spore formation:spo0A, spo0A, spo0E, spo0F, and spo0H. A mutation in any one of these regulatory genes severely restricted transcription of the 0.4 kb sequence. Transcription of the ctc gene was also turned on by nutrient deprivation, but this RNA synthesis was not impaired in spo0 mutants. Although not under spo0 control, the ctc gene probably corresponds to a locus, spoVC, whose product is required at a late stage of sporulation. Finally, the veg gene was actively transcribed both in growing cells and in nutrient-deprived cells. Like ctc RNA synthesis, transcription of the veg gene was not dependent upon the spo0 gene products. We propose that the spo0A, spo0B, spo0E, spo0F, and spo0H gene products are components of a pathway(s) that senses nutrient deprivation in B. subtilis and translates this environmental signal into the transcriptional activation of a subset of developmental genes.

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Selected References

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  1. Brehm S. P., Le Hegarat F., Hoch J. A. Deoxyribonucleic acid-binding proteins in vegetative Bacillus subtilis: alterations caused by stage O sporulation mutations. J Bacteriol. 1975 Nov;124(2):977–984. doi: 10.1128/jb.124.2.977-984.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Copeland J. C., Marmur J. Identification of conserved genetic functions in Bacillus by use of temperature-sensitive mutants. Bacteriol Rev. 1968 Dec;32(4 Pt 1):302–312. [PMC free article] [PubMed] [Google Scholar]
  3. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  4. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Haldenwang W. G., Banner C. D., Ollington J. F., Losick R., Hoch J. A., O'Connor M. B., Sonenshein A. L. Mapping a cloned gene under sporulation control by inserttion of a drug resistance marker into the Bacillus subtilis chromosome. J Bacteriol. 1980 Apr;142(1):90–98. doi: 10.1128/jb.142.1.90-98.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Haldenwang W. G., Lang N., Losick R. A sporulation-induced sigma-like regulatory protein from B. subtilis. Cell. 1981 Feb;23(2):615–624. doi: 10.1016/0092-8674(81)90157-4. [DOI] [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. Haldenwang W. G., Losick R. Novel RNA polymerase sigma factor from Bacillus subtilis. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7000–7004. doi: 10.1073/pnas.77.12.7000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hayward G. S. Gel electrophoretic separation of the complementary strands of bacteriophage DNA. Virology. 1972 Jul;49(1):342–344. doi: 10.1016/s0042-6822(72)80042-4. [DOI] [PubMed] [Google Scholar]
  11. Hsieh T., Wang J. C. Physicochomecial studies on interactions between DNA and RNA polymerase. Isolation and mapping of a T7 DNA fragment containing the early promoters for Escherichia coli RNA polymerase. Biochemistry. 1976 Dec 28;15(26):5776–5783. doi: 10.1021/bi00671a014. [DOI] [PubMed] [Google Scholar]
  12. Hutchison K. W., Halvorson H. O. Cloning of randomly sheared DNA fragments from a phi 105 lysogen of Bacillus subtilis identification of prophage-containing clones. Gene. 1980 Feb;8(3):267–278. doi: 10.1016/0378-1119(80)90004-9. [DOI] [PubMed] [Google Scholar]
  13. Linn T., Losick R. The program of protein synthesis during sporulation in Bacillus subtilis. Cell. 1976 May;8(1):103–114. doi: 10.1016/0092-8674(76)90191-4. [DOI] [PubMed] [Google Scholar]
  14. Moran C. P., Jr, Losick R., Sonenshein A. L. Identification of a sporulation locus in cloned Bacillus subtilis deoxyribonucleic acid. J Bacteriol. 1980 Apr;142(1):331–334. doi: 10.1128/jb.142.1.331-334.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakayama T., Williamson V., Burtis K., Doi R. H. Purification and properties of two RNA polymerases from sporulating cells of Bacillus subtilis. Eur J Biochem. 1978 Jul 17;88(1):155–164. doi: 10.1111/j.1432-1033.1978.tb12433.x. [DOI] [PubMed] [Google Scholar]
  16. Ollington J. F., Losick R. A cloned gene that is turned on at an intermediate stage of spore formation in Bacillus subtilis. J Bacteriol. 1981 Aug;147(2):443–451. doi: 10.1128/jb.147.2.443-451.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pero J., Hannett N. M., Talkington C. Restriction cleavage map of SP01 DNA: general location of early, middle, and late genes. J Virol. 1979 Jul;31(1):156–171. doi: 10.1128/jvi.31.1.156-171.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Segall J., Losick R. Cloned Bacillus subtilis DNA containing a gene that is activated early during sporulation. Cell. 1977 Aug;11(4):751–761. doi: 10.1016/0092-8674(77)90289-6. [DOI] [PubMed] [Google Scholar]
  22. Shorenstein R. G., Losick R. Comparative size and properties of the sigma subunits of ribonucleic acid polymerase from Bacillus subtilis and Escherichia coli. J Biol Chem. 1973 Sep 10;248(17):6170–6173. [PubMed] [Google Scholar]
  23. Smith H. O., Birnstiel M. L. A simple method for DNA restriction site mapping. Nucleic Acids Res. 1976 Sep;3(9):2387–2398. doi: 10.1093/nar/3.9.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sonenshein A. L., Roscoe D. H. The course of phage phi-e infection in sporulating cells of Bacillus subtilis strain 3610. Virology. 1969 Oct;39(2):265–275. doi: 10.1016/0042-6822(69)90047-6. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Talkington C., Pero J. Promoter recognition by phage SP01-modified RNA polymerase. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1185–1189. doi: 10.1073/pnas.75.3.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Young M. Use of temperature-sensitive mutants to study gene expression during sporulation in Bacillus subtilis. J Bacteriol. 1976 May;126(2):928–936. doi: 10.1128/jb.126.2.928-936.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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