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J. Bacteriol. doi:10.1128/JB.00477-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Role of Dipicolinic Acid in the Germination, Stability and Viability of Spores of Bacillus subtilis

Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, CT 06030-3305; and Department of Physics, East Carolina University, Greenville, NC 27858-4353

^* To whom correspondence should be addressed. Email: setlow{at}nso2.uchc.edu.

	Abstract

Spores of Bacillus subtilis spoVF strains that cannot synthesize dipicolinic acid (DPA) but take it up during sporulation were prepared in medium with various DPA concentrations and the germination and viability of these spores were measured as well as the DPA content in individual spores. Levels of some other small molecules were also measured in DPA-less spores. These studies have allowed the following conclusions. 1) Spores with no or low DPA levels that lack either the cortex-lytic enzyme (CLE) SleB or the receptors that respond to nutrient germinants could be isolated, but were unstable and spontaneously initiated early steps in spore germination. 2) Spores that lacked SleB and nutrient germinant receptors and also had low DPA levels were more stable. 3) Spontaneous germination of spores with no or low DPA levels was at least in part via activation of SleB. 4) The other redundant CLE, CwlJ, was activated only by the release of high levels of DPA from spores. 5) Low levels of DPA were sufficient for the viability of spores that lacked most /-type small, acid-soluble spore proteins. 6) DPA levels accumulated in spores prepared in low DPA-containing media varied greatly between individual spores, in contrast to more homogeneous DPA levels in individual spores made in media with high DPA concentrations. 7) At least the great majority of spores of several spoVF strains that contained no DPA also lacked other major spore small molecules, and had gone through some of the early reactions in spore germination.

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