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产气荚膜梭菌中受VirR/VirS系统调控的毒力基因的蛋白质组和转录组分析

Proteome and transcriptome analysis of the virulence genes regulated by the VirR/VirS system in Clostridium perfringens.

作者信息

Shimizu Takeshi, Shima Kensuke, Yoshino Ken-ichi, Yonezawa Kazuyoshi, Shimizu Tohru, Hayashi Hideo

机构信息

Department of Microbiology, Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Ten-nohdai, Tsukuba 305-8575, Japan.

出版信息

J Bacteriol. 2002 May;184(10):2587-94. doi: 10.1128/JB.184.10.2587-2594.2002.

Abstract

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.

摘要

利用野生型和virR突变体培养上清液的二维凝胶电泳,对产气荚膜梭菌中受双组分系统VirR/VirS调控的蛋白质进行了分析。基于基质辅助激光解吸电离飞行时间质谱,鉴定出7种正调控蛋白和8种负调控蛋白。转录组分析证实,15种蛋白中的7种在转录水平上受VirR/VirS系统调控,但其余蛋白在翻译后和分泌水平上由VirR/VirS导向的蛋白酶进行修饰。我们从培养上清液中纯化并鉴定了VirR/VirS导向的蛋白酶,确定其为一种梭菌蛋白酶。由于这种蛋白水解活性受到亮抑蛋白酶肽和抗蛋白酶的强烈抑制,因此得出结论,该蛋白酶是产气荚膜梭菌半胱氨酸蛋白酶家族的一员。

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本文引用的文献

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Complete genome sequence of Clostridium perfringens, an anaerobic flesh-eater.产气荚膜梭菌(一种厌氧性食肉菌)的全基因组序列
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