σ(E) is a mother cell-specific transcription factor of sporulating Bacillus subtilis that is derived from an inactive precursor protein (pro- σ(E). To examine the process that prevents σ(E) activity from developing in the forespore, we fused the σ(E) structural gene (sigE) to forespore- specific promoters (P(dacF) and P(spoIIIG), placed these fusions at sites on the B. subtilis chromosome which translocate into the forespore either early or late, and used Western blot analysis to monitor SigE accumulation and pro- σ(E) processing. sigE alleles, placed at sites which entered the forespore early, were found to generate more protein product than the same fusion placed at a late entering site. SigE accumulation and processing in the forespore were enhanced by null mutations in spoIIIE, a gene whose product is essential for translocation of the distal portion of the B. subtilis chromosome into the forespore. In other experiments, a chimera of pro-σ(E) and green fluorescence protein, previously shown to be unprocessed if it is synthesized within the forespore, was found to be processed in this compartment if coexpressed with the gene for the pro-σ(E)-processing enzyme, SpoIIGA. The need for spoIIGA coexpression is obviated in the absence of SpoIIIE. We interpret these results as evidence that selective degradation of both SigE and SpoIIGA prevent mature σ(E) from accumulating in the forespore compartment of wild-type B. subtilis. Presumably, a gene(s) located at a site that is distal to the origin of chromosome transfer is responsible for this phenomenon when it is translocated and expressed in the forespore.
ASJC Scopus subject areas
- Molecular Biology