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Journal of Bacteriology, July 2008, p. 4798-4807, Vol. 190, No. 14
0021-9193/08/$08.00+0     doi:10.1128/JB.00477-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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

Anil Magge,¹ Amanda C. Granger,^1, Paul G. Wahome,¹ Barbara Setlow,¹ Venkata R. Vepachedu,^1, Charles A. Loshon,¹ Lixin Peng,² De Chen,² Yong-qing Li,² and Peter Setlow^1*

Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3305,¹ Department of Physics, East Carolina University, Greenville, North Carolina 27858-4353²

Received 8 April 2008/ Accepted 1 May 2008

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 as well as the DPA content in individual spores were measured. Levels of some other small molecules in DPA-less spores were also measured. These studies have allowed the following conclusions. (i) Spores with no DPA 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. (ii) Spores that lacked SleB and nutrient germinant receptors and also had low DPA levels were more stable. (iii) Spontaneous germination of spores with no DPA or low DPA levels was at least in part via activation of SleB. (iv) The other redundant CLE, CwlJ, was activated only by the release of high levels of DPA from spores. (v) Low levels of DPA were sufficient for the viability of spores that lacked most /β-type small, acid-soluble spore proteins. (vi) DPA levels accumulated in spores prepared in low-DPA-containing media varied greatly between individual spores, in contrast to the presence of more homogeneous DPA levels in individual spores made in media with high DPA concentrations. (vii) 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.

Published ahead of print on 9 May 2008.

Present address: Middlebury College, Middlebury, VT.

Present address: Marine Biological Laboratory, Woods Hole, MA.

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