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副溶血性弧菌中CalR对cpsQ-mfpABC和mfpABC的转录调控

Transcriptional regulation of cpsQ-mfpABC and mfpABC by CalR in Vibrio parahaemolyticus.

作者信息

Gao He, Zhang Lingyu, Osei-Adjei George, Yang Wenhui, Zhou Dongsheng, Huang Xinxiang, Yang Huiying, Yin Zhe, Zhang Yiquan

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing, China.

School of Medicine, Jiangsu University, Zhenjiang, Jiangsu, China.

出版信息

Microbiologyopen. 2017 Aug;6(4). doi: 10.1002/mbo3.470. Epub 2017 Mar 20.

Abstract

The cpsQ-mfpABC locus is transcribed as two operons, i.e., cpsQ-mfpABC and mfpABC, in Vibrio parahaemolyticus, and both of them are all required for biofilm formation. CalR belongs to the LysR-type transcriptional regulator family, and was originally identified as a repressor of the swarming motility and T3SS1 genes expression in V. parahaemolyticus. In the present work, a combination of qRT-PCR, primer extension, LacZ fusion expression, electrophoretic mobility shift assay, and DNase I footprinting assays were employed to elucidate the regulatory mechanisms of cpsQ-mfpABC and mfpABC by CalR. One transcription start site for each operon was detected and their activities were activated by CalR. His-CalR protected two DNA regions upstream of mfpABC against DNase I digestion, but no binding sites were detected in the promoter region of cpsQ-mfpABC, suggesting a direct and an indirect regulatory manner for mfpABC and cpsQ-mfpABC transcription by CalR, respectively. Collectively, the results presented here confirmed a new physiological role for CalR that acts as an activator for cpsQ-mfpABC and mfpABC transcription.

摘要

在副溶血性弧菌中,cpsQ-mfpABC基因座被转录为两个操纵子,即cpsQ-mfpABC和mfpABC,它们都是生物膜形成所必需的。CalR属于LysR型转录调节因子家族,最初被鉴定为副溶血性弧菌群体运动和T3SS1基因表达的阻遏物。在本研究中,采用qRT-PCR、引物延伸、LacZ融合表达、电泳迁移率变动分析和DNase I足迹分析等方法,阐明CalR对cpsQ-mfpABC和mfpABC的调控机制。检测到每个操纵子的一个转录起始位点,其活性被CalR激活。His-CalR保护mfpABC上游的两个DNA区域免受DNase I消化,但在cpsQ-mfpABC的启动子区域未检测到结合位点,这表明CalR分别对mfpABC和cpsQ-mfpABC转录采取直接和间接的调控方式。总的来说,这里给出的结果证实了CalR作为cpsQ-mfpABC和mfpABC转录激活剂的新生理作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef1a/5552913/cb89ea7ee5b3/MBO3-6-na-g001.jpg

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