Evidence is presented that a sporulationessential or factor of Bacillus subtilis, a29, is synthesized as an inactive precursor (P3') and that its activation occurs by a developmentally regulated cleavage of 29 amino acids from the p31 amino terminus. A pulse-chase experiment demonstrated that r29 was derived from a preexisting protein, with appearance of radioactively labeled ar29 paralleling the disappearance of labeled P31. The disappearance of pulse-labeled P31 did not occur when the experiment was done with a B. subtilis strain carrying a mutation in a locus (spollE) GBq) to 25 uCi/ml; then either the cultures were harvested or the label was "chased" by the addition of a 103-fold excess of unlabeled methionine with continued incubation prior to harvesting. Cell extracts were prepared as described (4) and treated overnight at 4°C with anti-o-29 monoclonal antibody (4) in a 0.5-ml reaction mixture containing 10 mM Tris (pH 7.8), 10 mM EDTA, 0.3 mg ofphenylmethylsulfonyl fluoride, 0.1% NaDodSO4, 0.2% deoxycholate, 1% Triton X-100, a concentrated extract from 10 ml of B. subtilis culture, and 20,ug of antibody. Extracts then were incubated in the same reaction buffer with rabbit anti-mouse immunoglobulin for 4 hr, followed by incubation with washed Staphylococcus aureus cells (11)
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