Proteome and transcriptome analysis of the virulence genes regulated by the VirR/VirS system in Clostridium perfringens
Proteome and transcriptome analysis of the virulence genes regulated by the VirR/VirS system in Clostridium perfringens
复制标题
DOI：
10.1128/jb.184.10.2587-2594.2002
复制
发表时间：
2002-05-01
影响因子：
3.2
通讯作者：
Hayashi, H
中科院分区：
文献类型：
--
作者：
Shimizu, T;Shima, K;Hayashi, H
通过双向凝胶电泳分析产气荚膜梭菌野生型和virR突变体的培养上清液，分析了在双组分系统VirR/VirS控制下的蛋白质。基于基质辅助激光解吸电离-飞行时间/质谱，鉴定了7个正调控蛋白和8个负调控蛋白。转录组分析证实，15个蛋白质中有7个在转录水平上受VirR/VirS系统的调控，其余蛋白质在翻译后和分泌水平上受VirR/VirS系统的蛋白酶修饰。我们从培养上清中纯化并鉴定了VirR/VirS定向蛋白酶，并将其鉴定为一种梭菌蛋白酶。由于该蛋白酶的蛋白水解活性被亮抑酶肽和抗痛剂强烈抑制，因此推断该蛋白酶是C.产气荚膜杆菌
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.