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J Bacteriol. Oct 1995; 177(20): 5906–5911.
PMCID: PMC177417

Use of green fluorescent protein for visualization of cell-specific gene expression and subcellular protein localization during sporulation in Bacillus subtilis.

Abstract

We report the use of the green fluorescent protein (GFP) of Aequorea victoria to visualize cell-specific gene expression and protein subcellular localization during sporulation in Bacillus subtilis. Sporangia bearing the gene (gfp) for the green fluorescent protein fused to genes under the control of the sporulation transcription factor sigma F exhibited a forespore-specific pattern of fluorescence. Forespore-specific fluorescence could be detected with fusions to promoters that are utilized with low (csfB) and high (sspE-2G) efficiency by sigma F-containing RNA polymerase. Conversely, a mother cell-specific pattern of fluorescence was observed in sporangia bearing a transcriptional fusion of gfp to a spore coat protein gene (cotE) under the control of sigma E and an in-frame fusion to a regulatory gene (gerE) under the control of sigma K. An in-frame fusion of gfp to cotE demonstrated that GFP can also be used to visualize protein subcellular localization. In sporangia producing the CotE-GFP fusion protein, fluorescence was found to localize around the developing spore, and this localization was dependent upon SpoIVA, a morphogenetic protein known to determine proper localization of CotE.

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Selected References

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  • Brehm SP, Staal SP, Hoch JA. Phenotypes of pleiotropic-negative sporulation mutants of Bacillus subtilis. J Bacteriol. 1973 Sep;115(3):1063–1070. [PMC free article] [PubMed]
  • Bylund JE, Zhang L, Haines MA, Higgins ML, Piggot PJ. Analysis by fluorescence microscopy of the development of compartment-specific gene expression during sporulation of Bacillus subtilis. J Bacteriol. 1994 May;176(10):2898–2905. [PMC free article] [PubMed]
  • Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. [PubMed]
  • Cody CW, Prasher DC, Westler WM, Prendergast FG, Ward WW. Chemical structure of the hexapeptide chromophore of the Aequorea green-fluorescent protein. Biochemistry. 1993 Feb 9;32(5):1212–1218. [PubMed]
  • Cutting S, Mandelstam J. The nucleotide sequence and the transcription during sporulation of the gerE gene of Bacillus subtilis. J Gen Microbiol. 1986 Nov;132(11):3013–3024. [PubMed]
  • Cutting S, Oke V, Driks A, Losick R, Lu S, Kroos L. A forespore checkpoint for mother cell gene expression during development in B. subtilis. Cell. 1990 Jul 27;62(2):239–250. [PubMed]
  • Cutting S, Panzer S, Losick R. Regulatory studies on the promoter for a gene governing synthesis and assembly of the spore coat in Bacillus subtilis. J Mol Biol. 1989 May 20;207(2):393–404. [PubMed]
  • Driks A, Losick R. Compartmentalized expression of a gene under the control of sporulation transcription factor sigma E in Bacillus subtilis. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):9934–9938. [PMC free article] [PubMed]
  • Driks A, Roels S, Beall B, Moran CP, Jr, Losick R. Subcellular localization of proteins involved in the assembly of the spore coat of Bacillus subtilis. Genes Dev. 1994 Jan;8(2):234–244. [PubMed]
  • Errington J, Mandelstam J. Use of a lacZ gene fusion to determine the dependence pattern of sporulation operon spoIIA in spo mutants of Bacillus subtilis. J Gen Microbiol. 1986 Nov;132(11):2967–2976. [PubMed]
  • Fort P, Errington J. Nucleotide sequence and complementation analysis of a polycistronic sporulation operon, spoVA, in Bacillus subtilis. J Gen Microbiol. 1985 May;131(5):1091–1105. [PubMed]
  • Harry EJ, Pogliano K, Losick R. Use of immunofluorescence to visualize cell-specific gene expression during sporulation in Bacillus subtilis. J Bacteriol. 1995 Jun;177(12):3386–3393. [PMC free article] [PubMed]
  • Heim R, Prasher DC, Tsien RY. Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12501–12504. [PMC free article] [PubMed]
  • Karmazyn-Campelli C, Bonamy C, Savelli B, Stragier P. Tandem genes encoding sigma-factors for consecutive steps of development in Bacillus subtilis. Genes Dev. 1989 Feb;3(2):150–157. [PubMed]
  • Kroos L, Kunkel B, Losick R. Switch protein alters specificity of RNA polymerase containing a compartment-specific sigma factor. Science. 1989 Jan 27;243(4890):526–529. [PubMed]
  • Kunkel B, Losick R, Stragier P. The Bacillus subtilis gene for the development transcription factor sigma K is generated by excision of a dispensable DNA element containing a sporulation recombinase gene. Genes Dev. 1990 Apr;4(4):525–535. [PubMed]
  • Kunkel B, Sandman K, Panzer S, Youngman P, Losick R. The promoter for a sporulation gene in the spoIVC locus of Bacillus subtilis and its use in studies of temporal and spatial control of gene expression. J Bacteriol. 1988 Aug;170(8):3513–3522. [PMC free article] [PubMed]
  • Lewis PJ, Nwoguh CE, Barer MR, Harwood CR, Errington J. Use of digitized video microscopy with a fluorogenic enzyme substrate to demonstrate cell- and compartment-specific gene expression in Salmonella enteritidis and Bacillus subtilis. Mol Microbiol. 1994 Aug;13(4):655–662. [PubMed]
  • Lewis PJ, Partridge SR, Errington J. Sigma factors, asymmetry, and the determination of cell fate in Bacillus subtilis. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3849–3853. [PMC free article] [PubMed]
  • Losick R, Stragier P. Crisscross regulation of cell-type-specific gene expression during development in B. subtilis. Nature. 1992 Feb 13;355(6361):601–604. [PubMed]
  • Margolis P, Driks A, Losick R. Establishment of cell type by compartmentalized activation of a transcription factor. Science. 1991 Oct 25;254(5031):562–565. [PubMed]
  • Morin JG, Hastings JW. Energy transfer in a bioluminescent system. J Cell Physiol. 1971 Jun;77(3):313–318. [PubMed]
  • Morise H, Shimomura O, Johnson FH, Winant J. Intermolecular energy transfer in the bioluminescent system of Aequorea. Biochemistry. 1974 Jun 4;13(12):2656–2662. [PubMed]
  • Oke V, Losick R. Multilevel regulation of the sporulation transcription factor sigma K in Bacillus subtilis. J Bacteriol. 1993 Nov;175(22):7341–7347. [PMC free article] [PubMed]
  • Partridge SR, Errington J. The importance of morphological events and intercellular interactions in the regulation of prespore-specific gene expression during sporulation in Bacillus subtilis. Mol Microbiol. 1993 May;8(5):945–955. [PubMed]
  • Piggot PJ, Coote JG. Genetic aspects of bacterial endospore formation. Bacteriol Rev. 1976 Dec;40(4):908–962. [PMC free article] [PubMed]
  • Resnekov O, Driks A, Losick R. Identification and characterization of sporulation gene spoVS from Bacillus subtilis. J Bacteriol. 1995 Oct;177(19):5628–5635. [PMC free article] [PubMed]
  • Roels S, Driks A, Losick R. Characterization of spoIVA, a sporulation gene involved in coat morphogenesis in Bacillus subtilis. J Bacteriol. 1992 Jan;174(2):575–585. [PMC free article] [PubMed]
  • Setlow B, Magill N, Febbroriello P, Nakhimovsky L, Koppel DE, Setlow P. Condensation of the forespore nucleoid early in sporulation of Bacillus species. J Bacteriol. 1991 Oct;173(19):6270–6278. [PMC free article] [PubMed]
  • Stevens CM, Daniel R, Illing N, Errington J. Characterization of a sporulation gene, spoIVA, involved in spore coat morphogenesis in Bacillus subtilis. J Bacteriol. 1992 Jan;174(2):586–594. [PMC free article] [PubMed]
  • Sun D, Fajardo-Cavazos P, Sussman MD, Tovar-Rojo F, Cabrera-Martinez RM, Setlow P. Effect of chromosome location of Bacillus subtilis forespore genes on their spo gene dependence and transcription by E sigma F: identification of features of good E sigma F-dependent promoters. J Bacteriol. 1991 Dec;173(24):7867–7874. [PMC free article] [PubMed]
  • Sun DX, Stragier P, Setlow P. Identification of a new sigma-factor involved in compartmentalized gene expression during sporulation of Bacillus subtilis. Genes Dev. 1989 Feb;3(2):141–149. [PubMed]
  • Wang S, Hazelrigg T. Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis. Nature. 1994 Jun 2;369(6479):400–403. [PubMed]
  • Youngman P, Perkins JB, Losick R. Construction of a cloning site near one end of Tn917 into which foreign DNA may be inserted without affecting transposition in Bacillus subtilis or expression of the transposon-borne erm gene. Plasmid. 1984 Jul;12(1):1–9. [PubMed]
  • Zheng L, Halberg R, Roels S, Ichikawa H, Kroos L, Losick R. Sporulation regulatory protein GerE from Bacillus subtilis binds to and can activate or repress transcription from promoters for mother-cell-specific genes. J Mol Biol. 1992 Aug 20;226(4):1037–1050. [PubMed]

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