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PnpM,一种 LysR 型转录调节因子,激活了嗜麦芽窄食单胞菌 WBC-3 菌株中对硝基苯酚降解的对苯二酚途径。

PnpM, a LysR-Type Transcriptional Regulator Activates the Hydroquinone Pathway in -Nitrophenol Degradation in sp. Strain WBC-3.

作者信息

Wang Jin-Pei, Zhang Wen-Mao, Chao Hong-Jun, Zhou Ning-Yi

机构信息

Wuhan Institute of Virology, Chinese Academy of SciencesWuhan, China.

University of Chinese Academy of SciencesBeijing, China.

出版信息

Front Microbiol. 2017 Sep 14;8:1714. doi: 10.3389/fmicb.2017.01714. eCollection 2017.

Abstract

A LysR-type transcriptional regulator (LTTR), PnpR, has previously been shown to activate the transcription of operons , and for -nitrophenol (PNP) degradation in sp. strain WBC-3. Further preliminary evidence suggested the possible presence of an LTTR additional binding site in the promoter region of . In this study, an additional LTTR PnpM, which shows 44% homology to PnpR, was determined to activate the expression of . Interestingly, a -deleted WBC-3 strain was unable to grow on PNP but accumulating hydroquinone (HQ), which is the catabolic product from PNP degradation by PnpAB and the substrate for PnpCD. Through electrophoretic mobility shift assays (EMSAs) and promoter activity detection, only PnpR was involved in the activation of and , but both PnpR and PnpM were involved in the activation of . DNase I footprinting analysis suggested that PnpR and PnpM shared the same DNA-binding regions of 27 bp in the promoter. In the presence of PNP, the protection region increased to 39 bp by PnpR and to 38 bp by PnpM. Our data suggested that both PnpR and PnpM were involved in activating expression, in which PNP rather than the substrate hydroquinone for PnpCD is the inducer. Thus, during the PNP catabolism in sp. strain WBC-3, and operons for the initial two reactions were controlled by PnpR, while the third operon () for HQ degradation was activated by PnpM and PnpR. This study builds upon our previous findings and shows that two LTTRs PnpR and PnpM are involved in the transcriptional activation of these three catabolic operons. Specifically, our identification that an LTTR, PnpM, regulates expression provides new insights in an intriguing regulation system of PNP catabolism that is controlled by two regulators.

摘要

先前已证明,一种LysR型转录调节因子(LTTR)PnpR可激活操纵子的转录,并参与嗜麦芽窄食单胞菌WBC - 3菌株中对硝基苯酚(PNP)的降解。进一步的初步证据表明,在pnpCDE操纵子的启动子区域可能存在一个LTTR额外结合位点。在本研究中,确定了另一种与PnpR具有44%同源性的LTTR PnpM可激活pnpCDE的表达。有趣的是,一个pnpAB缺失的WBC - 3菌株无法在PNP上生长,但会积累对苯二酚(HQ),对苯二酚是PnpAB降解PNP的分解代谢产物以及PnpCD的底物。通过电泳迁移率变动分析(EMSA)和启动子活性检测,只有PnpR参与了pnpAB和pnpC的激活,但PnpR和PnpM都参与了pnpCDE的激活。DNase I足迹分析表明,PnpR和PnpM在pnpCDE启动子中共享27 bp的相同DNA结合区域。在存在PNP的情况下,PnpR使保护区域增加到39 bp,PnpM使其增加到38 bp。我们的数据表明,PnpR和PnpM都参与激活pnpCDE表达,其中PNP而非PnpCD的底物对苯二酚是诱导剂。因此,在嗜麦芽窄食单胞菌WBC - 3菌株的PNP分解代谢过程中,前两个反应的pnpAB和pnpC操纵子由PnpR控制,而HQ降解的第三个操纵子(pnpCDE)由PnpM和PnpR激活。本研究基于我们之前的发现,表明两个LTTR PnpR和PnpM参与了这三个分解代谢操纵子的转录激活。具体而言,我们鉴定出一种LTTR,即PnpM,可调节pnpCDE表达,这为一个由两个调节因子控制的有趣的PNP分解代谢调节系统提供了新见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7c4d/5603801/c494c657dd81/fmicb-08-01714-g0001.jpg

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