Analysis of the Transcriptional Regulator GlpR, Promoter Elements, and Posttranscriptional Processing Involved in Fructose-Induced Activation of the Phosphoenolpyruvate-Dependent Sugar Phosphotransferase System in Haloferax mediterranei----中国科学院微生物研究所

科研成果

论文题目:	Analysis of the Transcriptional Regulator GlpR, Promoter Elements, and Posttranscriptional Processing Involved in Fructose-Induced Activation of the Phosphoenolpyruvate-Dependent Sugar Phosphotransferase System in Haloferax mediterranei
作者:	Cai Lei, Cai Shuangfeng, Zhao Dahe, Wu Jinhua, Wang Lei, Liu Xiaoqing, Li Ming, Hou Jing, Zhou Jian, Liu Jingfang, Han Jing, Xiang Hua*
联系作者:	Xiang Hua*
刊物名称:	Applied and Environmental Microbiology
期:	80
卷:	4
页:	1430-1440
年份:	2014
影响因子:	4.406
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摘要:	Among all known archaeal strains, the phosphoenolpyruvate-dependent phosphotransferase system (PTS) for fructose utilization is used primarily by haloarchaea, which thrive in hypersaline environments, whereas the molecular details of the regulation of the archaeal PTS under fructose induction remain unclear. In this study, we present a comprehensive examination of the regulatory mechanism of the fructose PTS in the haloarchaeon Haloferax mediterranei. With gene knockout and complementation, microarray analysis, and chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR), we revealed that GlpR is the indispensable activator, which specifically binds to the PTS promoter (PPTS) during fructose induction. Further promoter-scanning mutation indicated that three sites located upstream of the H. mediterranei PPTS, which are conserved in most haloarchaeal P(PTS)s, are involved in this induction. Interestingly, two PTS transcripts (named T-8 and T-17) with different lengths of 5' untranslated region (UTR) were observed, and promoter or 5' UTR swap experiments indicated that the shorter 5' UTR was most likely generated from the longer one. Notably, the translation efficiency of the transcript with this shorter 5' UTR was significantly higher and the ratio of T-8 (with the shorter 5' UTR) to T-17 increased during fructose induction, implying that a posttranscriptional mechanism is also involved in PTS activation. With these insights into the molecular regulation of the haloarchaeal PTS, we have proposed a working model for haloarchaea in response to environmental fructose.