Transcriptional regulation of cpsQ-mfpABC and mfpABC by CalR in Vibrio parahaemolyticus.
Transcriptional regulation of cpsQ-mfpABC and mfpABC by CalR in Vibrio parahaemolyticus.
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副溶血弧菌中 CalR 对 cpsQ-mfpABC 和 mfpABC 的转录调节。
DOI：
10.1002/mbo3.470
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发表时间：
2017-08
期刊：
影响因子：
3.4
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Gao H;Zhang L;Osei-Adjei G;Yang W;Zhou D;Huang X;Yang H;Yin Z;Zhang Y
cpsQ-mfpABC基因座转录为两个操纵子，即，cpsQ-mfpABC和mfpABC，在副溶血性弧菌中，并且它们都是生物膜形成所需的。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中群集运动和T3 SS 1基因表达的阻遏物。在本工作中，采用qRT-PCR、引物延伸、LacZ融合表达、电泳迁移率变动分析和DNA酶I足迹分析的组合来阐明CalR对cpsQ-mfpABC和mfpABC的调控机制。每个操纵子都有一个转录起始位点，它们的活性都被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的启动子区域中未检测到结合位点，这表明CalR分别对mfpABC和cpsQ-mfpABC的转录进行直接和间接调控。总的来说，这里呈现的结果证实了CalR作为cpsQ-mfpABC和mfpABC转录的激活剂的新生理作用。
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.
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DOI：
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发表时间：
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