The sporulation-specific SpoVIF (YjcC) protein of Bacillus subtilis is essential for the development of heat-resistant spores. The GerE protein, the smallest member of the LuxR-FixJ family, contains a helix–turn–helix (HTH) motif and is involved in the expression of various sporulation-specific genes. In this study, the gene expression and protein composition of sporulating spoVIF-negative cells were analysed. CgeA, CotG and CotS, which are GerE-dependent coat proteins, were not expressed in the spoVIF-negative cells. Northern blotting showed that SpoVIF regulated the transcription of cgeA, cotG and cotS in a manner similar to that of GerE. In spoVIF-negative cells, gerE mRNA was transcribed normally, but immunoblot analysis using anti-GerE antiserum showed that the quantity of GerE protein was considerably less than that in wild-type controls. Using GFP (green fluorescent protein) fusion proteins, the localization of SpoVIF and GerE was observed by fluorescence microscopy. SpoVIF-GFP was detectable in the mother cell compartment, as was GerE-GFP. These results suggest that SpoVIF directly or indirectly controls the function of the GerE protein, and that SpoVIF is required for gene regulation during the latter stages of sporulation.
BauerT., LittleS., StoverA. G., DriksA.1999; Functional regions of the Bacillus subtilis spore coat morphogenetic protein CotE. J Bacteriol 181:7043–7051
CuttingS. M., Vander HornP. B.1990; Genetic analysis. In Molecular Biological Methods for Bacillus pp. 27–64Edited byHarwoodC. R., CuttingS. M. Chichester: Wiley;
DriksA.2001; Proteins of the spore core and coat. In Bacillus subtilis and its Closest Relatives from Genes to Cells pp. 527–535Edited bySonensheinA. L., HochJ. A., LosickR. Washington, DC: American Society for Microbiology;
DriksA., RoelsS., BeallB., MoranC. P.Jr, LosickR.1994; Subcellular localization of proteins involved in the assembly of the spore coat of Bacillus subtilis. Genes Dev 8:234–244[CrossRef]
HollandS. K., CuttingS., MandelstamJ.1987; The possible DNA-binding nature of the regulatory proteins, encoded by spoIID and gerE, involved in the sporulation of Bacillus subtilis. J Gen Microbiol 133:2381–2391
IgoM. M., LosickR.1986; Regulation of a promoter that is utilized by minor forms of RNA polymerase holoenzyme in Bacillus subtilis. J Mol Biol 191:615–624[CrossRef]
KahnD., DittaG.1991; Modular structure of FixJ: homology of the transcriptional activator domain with the −35 binding domain of sigma factors. Mol Microbiol 5:987–997[CrossRef]
KakeshitaH., TakamatsuH., AmikuraR., NakamuraK., WatabeK., YamaneK.2001; Effect of depletion of FtsY on spore morphology and the protein composition of the spore coat layer in Bacillus subtilis. FEMS Microbiol Lett 195:41–46[CrossRef]
KodamaT., TakamatsuH., AsaiK., KobayashiK., OgasawaraN., WatabeK.1999; The Bacillus subtilis yaaH gene is transcribed by SigE RNA polymerase during sporulation, and its product is involved in germination of spores. J Bacteriol 181:4584–4591
NakaneA., TakamatsuH., OguroA., SadaieY., NakamuraK., YamaneK.1995; Acquisition of azide-resistance by elevated SecA ATPase activity confers azide-resistance upon cell growth and protein translocation in Bacillus subtilis. Microbiology 141:113–121[CrossRef]
PiggotP. J., LosickR.2001; Sporulation genes and intercompartmental regulation. In Bacillus subtilis and its Closest Relatives from Genes To Cells pp. 483–517Edited bySonensheinA. L., HochJ. A., LosickR. Washington, DC: American Society for Microbiology;
PriceK. D., LosickR.1999; A four-dimensional view of assembly of a morphogenetic protein during sporulation in Bacillus subtilis. J Bacteriol 181:781–790
RoelsS., LosickR.1995; Adjacent and divergently oriented operons under the control of the sporulation regulatory protein GerE in Bacillus subtilis. J Bacteriol 177:6263–6275
SaccoM., RiccaE., LosickR., CuttingS.1995; An additional GerE-controlled gene encoding an abundant spore coat protein from Bacillus subtilis. J Bacteriol 177:372–377
SetlowP.1993; Spore structure proteins. In Bacillus subtilis and Other Gram-Positive Bacteria; Biochemistry, Physiology and Molecular Genetics pp. 801–809Edited bySonensheinA. L., HochJ. A., LosickR. Washington, DC: American Society for Microbiology;
TakamatsuH., ChikahiroY., KodamaT., KoideH., KozukaS., TochikuboK., WatabeK.1998; A spore coat protein, CotS, of Bacillus subtilis is synthesized under the regulation of σK and GerE during development and is located in the inner coat layer of spores. J Bacteriol 180:2968–2974
TakamatsuH., KodamaT., WatabeK.1999; Assembly of the CotSA coat protein into spores requires CotS in Bacillus subtilis.. FEMS Microbiol Lett 174:201–206[CrossRef]
WebbC. D., DecaturA., TelemanA., LosickR.1995; Use of green fluorescent protein for visualization of cell-specific gene expression and subcellular protein localization during sporulation in Bacillus subtilis. J Bacteriol 177:5906–5911
ZhengL., HalbergR., RoelsS., IchikawaH., KroosL., LosickR.1992; 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 226:1037–1050[CrossRef]