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Journal of Bacteriology, April 2004, p. 2413-2417, Vol. 186, No. 8
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.8.2413-2417.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Characterization of a Major Bacillus anthracis Spore Coat Protein and Its Role in Spore Inactivation

Ho-San Kim,1,{dagger} D. Sherman,2 F. Johnson,1 and A. I. Aronson1*

Department of Biological Sciences,1 Life Science Microscopy Facility, Purdue University, West Lafayette, Indiana 479072

Received 28 October 2003/ Accepted 30 December 2003

A major Bacillus anthracis spore coat protein of 13.4 kDa, designated Cot{alpha}, was found only in the Bacillus cereus group. A stable ca. 30-kDa dimer of this protein was also present in spore coat extracts. Cot{alpha}, which is encoded by a monocistronic gene, was first detected late in sporulation, consistent with a {sigma}K-regulated gene. On the basis of immunogold labeling, the protein is in the outer spore coat and absent from the exosporium. In addition, disruption of the gene encoding Cot{alpha} resulted in spores lacking a dark-staining outer spore coat in thin-section electron micrographs. The mutant spores were stable upon heating or storage, germinated at the same rate as the wild type, and were resistant to lysozyme. They were, however, more sensitive than the wild type to phenol, chloroform, and hypochlorite but more resistant to diethylpyrocarbonate. In all cases, resistance or sensitivity to these reagents was restored by introducing a clone of the cot{alpha} gene into the mutant. Since Cot{alpha} is an abundant outer spore coat protein of the B. cereus group with a prominent role in spore resistance and sensitivity, it is a promising target for the inactivation of B. anthracis spores.

* Corresponding author. Mailing address: Purdue University, Department of Biological Sciences, 1392 Lilly Hall of Life Sciences, West Lafayette, IN 47907. Phone: (765) 494-4992. Fax: (765) 494-0876. E-mail: aronson{at}purdue.edu.

{dagger} Present address: Department of Microbiology, Loyola University Medical School, Chicago, Ill.

Journal of Bacteriology, April 2004, p. 2413-2417, Vol. 186, No. 8
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.8.2413-2417.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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