Isolation of Bacillus subtilis genes transcribed in vitro and in vivo by a major sporulation-induced, DNA-dependent RNA polymerase - PubMed (original) (raw)
Isolation of Bacillus subtilis genes transcribed in vitro and in vivo by a major sporulation-induced, DNA-dependent RNA polymerase
G L Ray et al. J Bacteriol. 1986 May.
Abstract
As a means of determining the function of sigma 29, a sporulation-essential sigma factor, we have isolated and begun to characterize genes that require sigma 29 for their expression. RNA transcribed in vitro from total Bacillus subtilis DNA by using sigma 29-containing RNA polymerase (E-sigma-29) was hybridized to a bank of B. subtilis DNA fragments that had been cloned into bacteriophage lambda. Approximately 0.25% of the cloned B. subtilis DNA fragments displayed detectable hybridization with our RNA probe. Five DNA fragments that had strong in vitro template activity for E-sigma-29 were selected for further study. The DNA fragments which contained in vitro sigma 29 promoter activity encoded RNAs that were synthesized by B. subtilis during sporulation. Mutant B. subtilis that failed to synthesize sigma 29 (spoIIA, spoIIE) made less RNA that could hybridize to these cloned DNAs than did a mutant (spoIIC) which did synthesize sigma 29 but was blocked at a similar stage in development. A detailed analysis of several of the cloned DNAs demonstrated that they encoded RNAs that were transcribed from approximately the same start site in vivo that E-sigma-29 initiated transcription in vitro. These particular transcripts were present only during the period of sigma-29 abundance (2 to 4 h after the onset of sporulation) in sporulating cells which carried a wild-type allele of the sigma-29 structural gene (spoIIG). We conclude that the isolation procedure used in this study identified genes that are transcribed by E-sigma 29, not only in vitro but also in vivo. Preliminary characterization of the cloned genes indicate that they encoded multiple overlapping RNAs which were each synthesized at unique times during growth or sporulation. This result implies that sigma 29 does not activate a unique population of genes with a novel function in sporulation but rather that it has a temporal role in spore gene control, transcribing those genes required to be active during its period of abundance regardless of their specific function.
Similar articles
- Temporal regulation and forespore-specific expression of the spore photoproduct lyase gene by sigma-G RNA polymerase during Bacillus subtilis sporulation.
Pedraza-Reyes M, Gutiérrez-Corona F, Nicholson WL. Pedraza-Reyes M, et al. J Bacteriol. 1994 Jul;176(13):3983-91. doi: 10.1128/jb.176.13.3983-3991.1994. J Bacteriol. 1994. PMID: 8021181 Free PMC article. - A sporulation-induced sigma-like regulatory protein from B. subtilis.
Haldenwang WG, Lang N, Losick R. Haldenwang WG, et al. Cell. 1981 Feb;23(2):615-24. doi: 10.1016/0092-8674(81)90157-4. Cell. 1981. PMID: 6781761 - Novel RNA polymerase sigma factor from Bacillus subtilis.
Haldenwang WG, Losick R. Haldenwang WG, et al. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7000-4. doi: 10.1073/pnas.77.12.7000. Proc Natl Acad Sci U S A. 1980. PMID: 6784117 Free PMC article. - Expression of stage II genes during sporulation in Bacillus subtilis.
Moran CP Jr. Moran CP Jr. Res Microbiol. 1991 Sep-Oct;142(7-8):841-5. doi: 10.1016/0923-2508(91)90063-g. Res Microbiol. 1991. PMID: 1784821 Review. - Control of the cell-specificity of sigma F activity in Bacillus subtilis.
Errington J, Feucht A, Lewis PJ, Lord M, Magnin T, Najafi SM, Wilkinson JF, Yudkin MD. Errington J, et al. Philos Trans R Soc Lond B Biol Sci. 1996 Apr 29;351(1339):537-42. doi: 10.1098/rstb.1996.0052. Philos Trans R Soc Lond B Biol Sci. 1996. PMID: 8735276 Review.
Cited by
- ROMA: an in vitro approach to defining target genes for transcription regulators.
Maclellan SR, Eiamphungporn W, Helmann JD. Maclellan SR, et al. Methods. 2009 Jan;47(1):73-7. doi: 10.1016/j.ymeth.2008.10.009. Epub 2008 Oct 21. Methods. 2009. PMID: 18948201 Free PMC article. Review. - Evidence that the spoIIM gene of Bacillus subtilis is transcribed by RNA polymerase associated with sigma E.
Smith K, Youngman P. Smith K, et al. J Bacteriol. 1993 Jun;175(11):3618-27. doi: 10.1128/jb.175.11.3618-3627.1993. J Bacteriol. 1993. PMID: 8501065 Free PMC article. - The sigma factors of Bacillus subtilis.
Haldenwang WG. Haldenwang WG. Microbiol Rev. 1995 Mar;59(1):1-30. doi: 10.1128/mr.59.1.1-30.1995. Microbiol Rev. 1995. PMID: 7708009 Free PMC article. Review. - Characteristics of an RNA polymerase population isolated from Bacillus subtilis late in sporulation.
Cummings CW, Haldenwang WG. Cummings CW, et al. J Bacteriol. 1988 Dec;170(12):5863-9. doi: 10.1128/jb.170.12.5863-5869.1988. J Bacteriol. 1988. PMID: 3142858 Free PMC article. - Genetic analysis of RNA polymerase-promoter interaction during sporulation in bacillus subtilis.
Ray C, Tatti KM, Jones CH, Moran CP Jr. Ray C, et al. J Bacteriol. 1987 May;169(5):1807-11. doi: 10.1128/jb.169.5.1807-1811.1987. J Bacteriol. 1987. PMID: 3106319 Free PMC article.
References
- Proc Natl Acad Sci U S A. 1965 Sep;54(3):704-11 - PubMed
- Biochim Biophys Acta. 1963 Aug 20;72:619-29 - PubMed
- J Mol Biol. 1975 Nov 5;98(3):503-17 - PubMed
- Science. 1977 Apr 8;196(4286):180-2 - PubMed
- Cell. 1977 Nov;12(3):721-32 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials