Variety of Sporulation Phenotypes Resulting from Mutations in a Single Regulatory Locus, spoIIA, in Bacillus subtilis

Abstract

Closely linked mutations in either of the two putative genes of the sporulation locus spoIIA can affect, in quite diverse ways, spore incidence, the production of alkaline phosphatase and DNAase, and the stability of the cells in sporulation medium. It is concluded that the locus has a regulatory function affecting the activation or induction of at least two, and possibly more, sporulation-associated operons.

Medium-dependent sporulation resulting from a mutation in the spollAB gene of Bacillus subtilis

“…(1990) and the enzyme activity was determined from a 0.5 ml sample of cell suspension. Alkaline phosphatase (APase) activity was assayed as described by Errington & Mandelstam (1983). One unit of APase activity is the amount of enzyme that hydrolyses 1 nmol p-nitrophenyl phosphate in 1 min at 30 "C. Cloning spollAC mutations suppressing the effects of spollAB22.…”

Identification of sporulation genes by genome-wide analysis of the σ E regulon of Bacillus subtilis

“…Cells were harvested from 50 ml cultures 2 h after the initiation of sporulation, pelleted and frozen in liquid nitrogen. In parallel, a sample was taken and assayed for alkaline phosphatase activity as a means of checking s E activity (Errington & Mandelstam, 1983;Partridge & Errington, 1993). The cell pellets were thawed at 37 uC in 1 ml TE containing 25 mg lysozyme ml 21 for 5 min; 1 ml 150 mM NaCl, 0?1 % SDS, 10 mM EDTA, 10 mg Pronase ml 21 was then added, mixed gently and the mixture incubated for a further 5 min.…”

Establishment of cell‐specific transcription during sporulation in Bacillus subtilis

“…In these experiments, expression should be abolished in any mutant that fails to synthesize mature a^. Of particular value in this context are a series of mutations in the gene encoding (f (spollAC), which fall into two distinct phenotypic classes (Errington and Mandelstam, 1983), now known to differ in respect of either preventing pro-o^ processing (the 'classical' spollAC mutations) or allowing it to occur ('spoiiAC(Py mutations; see Illing and Errington, 1991a). The nature of the role of CT'^ in processing of pro-o^ has not been resolved (see below) but it is nevertheless clear that genes which, like spollD (Clarke et al, 1986), are expressed in spollAC(P) mutants but blocked in the other class of mutant, are likely to be transcribed from o^-dependent promoters.…”