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Journal of Bacteriology, August 2001, p. 4886-4893, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4886-4893.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genetic Requirements for Induction of Germination of Spores of Bacillus subtilis by Ca2+-Dipicolinate

Madan Paidhungat, Katerina Ragkousi, and Peter Setlow*

Department of Biochemistry, University of Connecticut Health Center, Farmington, Connecticut 06032

Received 28 March 2001/Accepted 30 May 2001

Dormant Bacillus subtilis spores can be induced to germinate by nutrients, as well as by nonmetabolizable chemicals, such as a 1:1 chelate of Ca2+ and dipicolinic acid (DPA). Nutrients bind receptors in the spore, and this binding triggers events in the spore core, including DPA excretion and rehydration, and also activates hydrolysis of the surrounding cortex through mechanisms that are largely unknown. As Ca2+-DPA does not require receptors to induce spore germination, we asked if this process utilizes other proteins, such as the putative cortex-lytic enzymes SleB and CwlJ, that are involved in nutrient-induced germination. We found that Ca2+-DPA triggers germination by first activating CwlJ-dependent cortex hydrolysis; this mechanism is different from nutrient-induced germination where cortex hydrolysis is not required for the early germination events in the spore core. Nevertheless, since nutrients can induce release of the spore's DPA before cortex hydrolysis, we examined if the DPA excreted from the core acts as a signal to activate CwlJ in the cortex. Indeed, endogenous DPA is required for nutrient-induced CwlJ activation and this requirement was partially remedied by exogenous Ca2+-DPA. Our findings thus define a mechanism for Ca2+-DPA-induced germination and also provide the first definitive evidence for a signaling pathway that activates cortex hydrolysis in response to nutrients.

* Corresponding author. Mailing address: Department of Biochemistry, MC 3305, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06032. Phone: (860) 679 2607. Fax: (860) 679 3408. E-mail: setlow{at}nso2.uchc.edu.

Journal of Bacteriology, August 2001, p. 4886-4893, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4886-4893.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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