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J Bacteriol, June 1998, p. 2968-2974, Vol. 180, No. 11
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Spore Coat Protein, CotS, of Bacillus subtilis Is Synthesized under the Regulation of sigma K and GerE during Development and Is Located in the Inner Coat Layer of Spores

Hiromu Takamatsu,1 Yukari Chikahiro,1,dagger Takeko Kodama,1 Hidekatsu Koide,2 Satoshi Kozuka,3 Kunio Tochikubo,3 and Kazuhito Watabe1,*

Faculty of Pharmaceutical Sciences, Setsunan University, Osaka,1 Meditopia Research Center, Tokyo,2 and Department of Microbiology, Nagoya City University Medical School, Nagoya,3 Japan

Received 23 October 1997/Accepted 31 March 1998

The spore coat of Bacillus subtilis has a unique morphology and consists of polypeptides of different sizes, whose synthesis and assembly are precisely regulated by a cascade of transcription factors and regulatory proteins. We examined the factors that regulate cotS gene expression and CotS assembly into the coat layer of B. subtilis by Northern blot and Western blot analysis. Transcription of cotS mRNA was not detected in sporulating cells of sigma K and gerE mutants by Northern blot analysis. By Western blot analysis using anti-CotS antibody, CotS was first detected in protein samples solubilized from wild-type cells at 5 h after the start of sporulation. CotS was not detected in the vegetative cells and spores of a gerE mutant or in the spores of mutants deficient in sigma E, sigma F, sigma G, or sigma K. CotS was detected in the sporangium but not in the spores of a cotE mutant. The sequence of the promoter region of cotS was similar to the consensus sequences for binding of sigma K and GerE. These results demonstrate that sigma K and GerE are required for cotS expression and that CotE is essential for the assembly of CotS in the coat. Immunoelectron microscopic observation using anti-CotS antibody revealed that CotS is located within the spore coat, in particular in the inner coats of dormant spores.

* Corresponding author. Mailing address: Faculty of Pharmaceutical Sciences, Setsunan University, Hirakata, Osaka 573-0101, Japan. Phone and fax: (81) 720 66 3112. E-mail: watabe{at}pharm.setsunan.ac.jp.

dagger Present address: Yakujyu Pharmacy, Yamato, Kanagawa, Japan.

J Bacteriol, June 1998, p. 2968-2974, Vol. 180, No. 11
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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