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[GroEL/GroES伴侣蛋白系统和Lon蛋白酶在大肠杆菌细胞中调节费氏弧菌lux基因表达的作用]

[Role of GroEL/GroES chaperonin system and Lon protease in regulation of expression Vibrio fischeri lux genes in Escherichia coli cells].

作者信息

Manukhov I V, Kotova V Iu, Zavil'genskiĭ G B

出版信息

Mol Biol (Mosk). 2006 Mar-Apr;40(2):277-83.

Abstract

It was shown that the chaperonin GroEL/GroES and protease Lon influence the expression of the Vibrio fischeri lux regulon in Escherichia coli cells: E. coli groE mutants bearing hybrid plasmid with the lux regulon were weakly luminescent; cells of the E. coli lon- comprising the entire lux regulon display very intense bioluminescence, with no lag period in the induction curve characteristic of lon+ strains. The luxR gene was cloned from the Vibrio fischeri genome in the pGEX-KG vector. It was shown that the active fusion protein GST-LuxR by affinity chromatography on glutathione-sucrose colony is purified only with proteins GroEL and Lon. The present results showed that the LuxR, transcriptional activator of the V. fischeri lux operon, really complexes with GroEL chaperonin and Lon protease. We suppose, that the GroEL/GroES chaperonin systems is required for the folding of LuxR into an active protein, and the LuxR is the target for the ATP-dependent serine Lon protease of E. coli.

摘要

结果表明,伴侣蛋白GroEL/GroES和蛋白酶Lon影响费氏弧菌lux操纵子在大肠杆菌细胞中的表达:携带lux操纵子杂交质粒的大肠杆菌groE突变体发光较弱;包含整个lux操纵子的大肠杆菌lon-细胞表现出非常强烈的生物发光,诱导曲线中没有lon+菌株特有的延迟期。从费氏弧菌基因组中克隆luxR基因到pGEX-KG载体中。结果表明,活性融合蛋白GST-LuxR通过谷胱甘肽-蔗糖柱亲和层析仅与蛋白质GroEL和Lon一起纯化。目前的结果表明,费氏弧菌lux操纵子的转录激活因子LuxR确实与伴侣蛋白GroEL和蛋白酶Lon形成复合物。我们推测,GroEL/GroES伴侣蛋白系统是LuxR折叠成活性蛋白所必需的,并且LuxR是大肠杆菌ATP依赖性丝氨酸Lon蛋白酶的作用靶点。

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