Alternative Translation Initiation Produces a Short Form of a Spore Coat Protein in Bacillus subtilis

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Abstract
During endospore formation in Bacillus subtilis, over two dozen polypeptides are localized to the developing spore and coordinately assembled into a thick multilayered structure called the spore coat. Assembly of the coat is initiated by the expression of morphogenetic proteins SpoIVA, CotE, and SpoVID. These morphogenetic proteins appear to guide the assembly of other proteins into the spore coat. For example, SpoVID forms a complex with the SafA protein, which is incorporated into the coat during the early stages of development. At least two forms of SafA are found in the mature spore coat: a full-length form and a shorter form (SafA-C 30 ) that begins with a methionine encoded by codon 164 of safA . In this study, we present evidence that the expression of SafA-C 30 arises from translation initiation at codon 164. We found only a single transcript driving expression of SafA. A stop codon engineered just upstream of a predicted ribosome-binding site near codon M164 abolished formation of full-length SafA, but not SafA-C 30 . The same effect was observed with an alanine substitution at codon 1 of SafA. Accumulation of SafA-C 30 was blocked by substitution of an alanine codon at codon 164, but not by a substitution at a nearby methionine at codon 161. We found that overproduction of SafA-C 30 interfered with the activation of late mother cell-specific transcription and caused a strong sporulation block.