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Journal of Bacteriology, May 2002, p. 2587-2594, Vol. 184, No. 10
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.10.2587-2594.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Proteome and Transcriptome Analysis of the Virulence Genes Regulated by the VirR/VirS System in Clostridium perfringens

Takeshi Shimizu,1 Kensuke Shima,1 Ken-ichi Yoshino,2 Kazuyoshi Yonezawa,2 Tohru Shimizu,1 and Hideo Hayashi1*

Department of Microbiology, Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Ten-nohdai, Tsukuba 305-8575,1 Biosignal Research Center, Kobe University, 1-1 Rokkodai Nada, Kobe 657-8501, Japan2

Received 26 November 2001/ Accepted 18 February 2002

The proteins under the control of the two-component system VirR/VirS in Clostridium perfringens were analyzed by using two-dimensional gel electrophoresis of the culture supernatant from the wild type and the virR mutant. Based on matrix-assisted laser desorption ionization-time of flight/mass spectrometry, seven positively regulated proteins and eight negatively regulated proteins were identified. Transcriptome analysis confirmed that 7 of the 15 proteins were regulated by the VirR/VirS system at the transcriptional level, but the remaining proteins were modified with a VirR/VirS-directed protease at the posttranslation and secretion levels. We purified and characterized the VirR/VirS-directed protease from the culture supernatant and identified it as a kind of clostripain. Because this proteolytic activity was strongly inhibited by leupeptin and antipain, it was concluded that this protease was a member of the family of cysteine proteases of C. perfringens.

* Corresponding author. Mailing address: Department of Microbiology, Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Ten-nohdai, Tsukuba, Japan 305-8575. Phone: 81-298-53-3354. Fax: 81-298-53-3354. E-mail: hihayash{at}md.tsukuba.ac.jp.

Journal of Bacteriology, May 2002, p. 2587-2594, Vol. 184, No. 10
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.10.2587-2594.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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