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创伤弧菌弹性蛋白酶基因通过两种不同类型的启动子以生长阶段依赖性方式进行差异表达。

Differential expression of Vibrio vulnificus elastase gene in a growth phase-dependent manner by two different types of promoters.

作者信息

Jeong H S, Jeong K C, Choi H K, Park K J, Lee K H, Rhee J H, Choi S H

机构信息

Department of Food Science and Technology, Department of Molecular Biotechnology, Institute of Biotechnology, Chonnam National University, Kwang-Ju 500-757, South Korea.

出版信息

J Biol Chem. 2001 Apr 27;276(17):13875-80. doi: 10.1074/jbc.M010567200. Epub 2001 Jan 31.

Abstract

Elastase activity of Vibrio vulnificus was highly dependent on growth phase, reached a maximum during the stationary phase, and was regulated at the level of transcription. The stationary phase production of elastase in crp or rpoS mutants, which were constructed by allelic exchanges, decreased about 3- and 10-fold, respectively. However, the promoter activity of vvpE encoding elastase was unaffected by those mutations in the log phase when analyzed using a vvpE-lux fusion. A primer extension analysis revealed that the transcription of vvpE begins at two different sites, consisting of putative promoter L (PL) and promoter S (PS). The PL activity was constitutive through the log and stationary phases, lower than the PS activity, and unaffected by the crp or rpoS mutations. The transcription of PS, induced only in the stationary phase, was dependent on RpoS. The mutation in crp reduced the activity of PS; however, the additional inactivation of crp did not influence the PS activity in the rpoS mutant, indicating that CRP exerted its effects through PS requiring RpoS. These results demonstrate that vvpE expression is differentially directed by PL and PS depending on the growth phase and elevated by RpoS and CRP in the stationary phase.

摘要

创伤弧菌的弹性蛋白酶活性高度依赖于生长阶段,在稳定期达到最大值,并在转录水平受到调控。通过等位基因交换构建的crp或rpoS突变体中,弹性蛋白酶在稳定期的产量分别下降了约3倍和10倍。然而,当使用vvpE-lux融合进行分析时,编码弹性蛋白酶的vvpE的启动子活性在对数期不受这些突变的影响。引物延伸分析表明,vvpE的转录起始于两个不同的位点,由假定的启动子L(PL)和启动子S(PS)组成。PL活性在对数期和稳定期都是组成型的,低于PS活性,并且不受crp或rpoS突变的影响。仅在稳定期诱导的PS转录依赖于RpoS。crp中的突变降低了PS的活性;然而,crp的额外失活并不影响rpoS突变体中的PS活性,这表明CRP通过需要RpoS的PS发挥其作用。这些结果表明,vvpE的表达根据生长阶段由PL和PS差异指导,并在稳定期由RpoS和CRP上调。

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