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Journal of Bacteriology, June 2006, p. 3740-3747, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.00212-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Role of Dipicolinic Acid in Resistance and Stability of Spores of Bacillus subtilis with or without DNA-Protective {alpha}/ß-Type Small Acid-Soluble Proteins

Barbara Setlow, Swaroopa Atluri, Ryan Kitchel, Kasia Koziol-Dube, and Peter Setlow*

Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3305

Received 8 February 2006/ Accepted 16 March 2006

Dipicolinic acid (DPA) comprises ~10% of the dry weight of spores of Bacillus species. Although DPA has long been implicated in spore resistance to wet heat and spore stability, definitive evidence on the role of this abundant molecule in spore properties has generally been lacking. Bacillus subtilis strain FB122 (sleB spoVF) produced very stable spores that lacked DPA, and sporulation of this strain with DPA yielded spores with nearly normal DPA levels. DPA-replete and DPA-less FB122 spores had similar levels of the DNA protective {alpha}/ß-type small acid-soluble spore proteins (SASP), but the DPA-less spores lacked SASP-{gamma}. The DPA-less FB122 spores exhibited similar UV resistance to the DPA-replete spores but had lower resistance to wet heat, dry heat, hydrogen peroxide, and desiccation. Neither wet heat nor hydrogen peroxide killed the DPA-less spores by DNA damage, but desiccation did. The inability to synthesize both DPA and most {alpha}/ß-type SASP in strain PS3664 (sspA sspB sleB spoVF) resulted in spores that lost viability during sporulation, at least in part due to DNA damage. DPA-less PS3664 spores were more sensitive to wet heat than either DPA-less FB122 spores or DPA-replete PS3664 spores, and the latter also retained viability during sporulation. These and previous results indicate that, in addition to {alpha}/ß-type SASP, DPA also is extremely important in spore resistance and stability and, further, that DPA has some specific role(s) in protecting spore DNA from damage. Specific roles for DPA in protecting spore DNA against damage may well have been a major driving force for the spore's accumulation of the high levels of this small molecule.

* Corresponding author. Mailing address: Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, CT 06030-3305. Phone: (860) 679-2607. Fax: (860) 679-3408. E-mail: setlow{at}nso2.uchc.edu.

Journal of Bacteriology, June 2006, p. 3740-3747, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.00212-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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