Protein Synthesis in the Isolated Forespores from Sporulating Cells of Bacillus subtilis

Developing forespores were isolated from Bacillus subtilis at different stages of sporulation and protein synthesis in the forespore compartment was examined. Pulse-labeling experiments indicated that ¹⁴C]phenylalanine was continuously incorporated into the sporangium throughout sporulation, and at t₅ (early stage V of sporulation) 58% of the radioactivity was located in the forespore compartment. Significantly high incorporation of ¹⁴C]phenylalanine was observed when the isolated forespores at t₅ were incubated with the corresponding mother-cell cytoplasmic fraction or an amino acid mixture. About 73% of the radioactivity incorporated into the isolated forespore at t₅ was found in the cytoplasmic fraction and 26% in the membranous fraction. Analysis by sodium dodecyl sulfate-gel electrophoresis showed that the ¹⁴C-labeled cytoplasmic protein had a molecular weight of about 20,000, and that a protein having the same molecular weight was present in the t₅ forespore as a slight protein band and also in the mature spore as a clear protein band. Gel electrophoresis also revealed that the ¹⁴C-labeled membranous-soluble protein (prepared by solubilization with detergents) had broad peaks with molecular weights of about 74,000, 33,000, 20,000, and 12,000.