Transcription of exsD is repressed directly by H-NS in Vibrio parahaemolyticus.
Vibrio parahaemolyticus is a leading cause of seafood-associated diarrhea and gastroenteritis. This bacteria expresses a major virulence determinant called T3SS1. Expression of T3SS1 is tightly regulated by the ExsA-ExsC-ExsD regulatory system. The transcription of exsA and probably exsC is repressed directly by the H-NS protein. In this study, the regulation of exsD by H-NS was investigated using quantitative RT-PCR, primer extension, LacZ fusion, electrophoretic mobility shift, and DNase I footprinting assays. The results showed that His-H-NS protected a single region from 61 bp to 174 bp upstream of exsD against DNase I digestion, and a transcription start site located at 105 bp upstream of exsD was detected and its activity was repressed by H-NS. Therefore, a single H-NS-dependent promoter was transcribed for exsD in V. parahaemolyticus. Thus, all three genes in the ExsA-ExsC-ExsD regulatory system of T3SS1 are directly repressed by H-NS in V. parahaemolyticus.
School of Medicine, Jiangsu University, Zhenjiang, 212013, Jiangsu, China.
Yiquan
Y
Zhang
School of Medicine, Jiangsu University, Zhenjiang, 212013, Jiangsu, China.
George
G
Osei-Adjei
School of Medicine, Jiangsu University, Zhenjiang, 212013, Jiangsu, China.
Bin
B
Ni
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100000, China; Department of Microbiology, Anhui Medical University, Hefei, 230032, Anhui, China.
Haihong
H
Fang
School of Medicine, Jiangsu University, Zhenjiang, 212013, Jiangsu, China.
Lingyu
L
Zhang
School of Medicine, Jiangsu University, Zhenjiang, 212013, Jiangsu, China.
Xin
X
Zhao
School of Medicine, Jiangsu University, Zhenjiang, 212013, Jiangsu, China.
Xinxiang
X
Huang
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100000, China.
Huiying
H
Yang
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100000, China. Electronic address: fionyoung@163.com.
Wenhui
W
Yang
Department of Pharmacy, Southwest Hospital, The Third Military Medical University, Chongqing, 400038, China. Electronic address: fengj_sun@163.com.
Fengjun
F
Sun
D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000378:metabolism*; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005809:Genes, Regulator; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014736:Vibrio parahaemolyticus / Q000235:genetics* / Q000378:metabolism*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C031908:H-NS protein, bacteria; D008565:Membrane Proteins; D001616:beta-Galactosidase
H-NS; T3SS1; Vibrio parahaemolyticus; exsD