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J. Bacteriol. doi:10.1128/JB.01353-06
Copyright (c) 2006, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

The timing of cotE expression affects Bacillus subtilis spore coat morphology but not lysozyme resistance

Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras, Portugal; Ernst-Moritz-Arndt-University, Medical Faculty, Laboratory for Functional Genomics, D-17487 Greifswald, Germany; Emory University School of Medicine, Department of Microbiology, Atlanta, GA 30322, USA

^* To whom correspondence should be addressed. Email: aoh{at}itqb.unl.pt.

	Abstract

The synthesis of structural components and morphogenetic factors required for the assembly of the Bacillus subtilis spore coat is governed by a mother cell-specific transcriptional cascade. The first two temporal classes of gene expression, which involve RNA polymerase sigma ^E factor and the ancillary regulators, GerR and SpoIIID, are deployed prior to engulfment of the prespore by the mother cell. The two last classes rely on ^K whose activation follows engulfment completion, and GerE. The cotE gene, codes for a morphogenetic protein essential for the assembly of the outer coat layer and spore resistance to lysozyme. cotE is expressed first from a ^E-dependent promoter and in a second stage, from a promoter that additionally requires SpoIIID, and that remains active under ^K control. CotE localizes prior to engulfment completion close to the surface of the developing spore, but formation of the outer coat is a late, ^K-controlled event. We have transplanted cotE to progressively later classes of mother cell gene expression. This created an early class of mutants, in which cotE is expressed prior to engulfment completion, and a late class in which expression of cotE follows the complete engulfment of the prespore. Mutants of the early class assemble a nearly normal outer coat structure, whereas mutants of the late class do not. Hence, the early expression of CotE is essential for outer coat assembly. Surprisingly however, all mutants were fully resistant to lysozyme. The results suggest that CotE has genetically separable functions in spore resistance to lysozyme, and spore outer coat assembly.

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