BACILLUS-SUBTILIS SPOVE GENE IS TRANSCRIBED BY SIGMA-E-ASSOCIATED RNA-POLYMERASE

Expression of the Bacillus subtilis sporulation gene spoVE was examined by runoff transcription assay with an RNA polymerase preparation obtained from vegetative and sporulating cells. Transcripts from tandem promoters (P1 and P2 promoters) located just upstream of the spoVE structure gene were detected. The transcription of spoVE initiated within an hour after the onset of sporulation and coincided with the presence of RNA polymerase associated with a 33-kDa protein. Amino acid sequence analysis of the 33-kDa protein revealed that it is a sigma factor, sigma(E). Reconstitution analysis of sigma(E) purified from the sporulating cell extracts and vegetative core RNA polymerase showed that sigma(E) recognizes the P2 promoter. SpoVE protein could not be synthesized in the transcription-translation coupled system prepared from vegetative cells (M. Okamoto, S. Fukui, and Y. Kobayashi, Agric. Biol. Chem. 49:1077-1082, 1985). However, addition of sigma(E)-associated RNA polymerase to the coupled system restored SpoVE protein synthesis. These results indicate that spoVE expression in sporulating cells is controlled essentially by sigma(E)-associated RNA polymerase.