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The wing of the ToxR winged helix-turn-helix domain is required for DNA binding and activation of toxT and ompU
- Source :
- PLoS ONE, Vol 14, Iss 9, p e0221936 (2019), PLoS ONE
- Publication Year :
- 2019
- Publisher :
- Public Library of Science (PLoS), 2019.
-
Abstract
- ToxR and TcpP, two winged helix-turn-helix (w-HTH) family transcription factors, co-activate expression of the toxT promoter in Vibrio cholerae. ToxT then directly regulates a number of genes required for virulence. In addition to co-activation of toxT, ToxR can directly activate the ompU promoter and repress the ompT promoter. Based on a previous study suggesting that certain wing residues of ToxR are preferentially involved in toxT co-activation compared to direct ompU activation, we employed alanine-scanning mutagenesis to determine which residues in the wing of ToxR are required for activation of each promoter. All of the ToxR wing residues tested that were critical for transcriptional activation of toxT and/or ompU were also critical for DNA binding. While some ToxR wing mutants had reduced interaction with TcpP, that reduced interaction did not correlate with a specific defect in toxT activation. Rather, such mutants also affected ompU activation and DNA binding. Based on these findings we conclude that the primary role of the wing of ToxR is to bind DNA, along with the DNA recognition helix of ToxR, and this function is required both for direct activation of ompU and co-activation of toxT.
- Subjects :
- Models, Molecular
Protein Extraction
Cell Membranes
Mutant
Helix-turn-helix
Plasma protein binding
Pathology and Laboratory Medicine
medicine.disease_cause
Biochemistry
Medicine and Health Sciences
Enzyme-Linked Immunoassays
Extraction Techniques
0303 health sciences
Mutation
Crystallography
Multidisciplinary
Chemistry
Physics
Chromosomes, Bacterial
Condensed Matter Physics
OmpT
Bacterial Pathogens
Cell biology
DNA-Binding Proteins
Medical Microbiology
Vibrio cholerae
Physical Sciences
Crystal Structure
Medicine
Chemical characterization
Cellular Structures and Organelles
Pathogens
Protein Binding
Research Article
Transcriptional Activation
animal structures
Science
DNA transcription
DNA binding assay
Microbiology
03 medical and health sciences
Bacterial Proteins
Protein Domains
Vibrio Cholerae
Genetics
Binding analysis
medicine
Solid State Physics
Adhesins, Bacterial
Immunoassays
Microbial Pathogens
Transcription factor
Helix-Turn-Helix Motifs
Vibrio
030304 developmental biology
Biology and life sciences
Bacteria
030306 microbiology
Mutagenesis
Organisms
Proteins
Membrane Proteins
DNA
Cell Biology
Outer Membrane Proteins
Research and analysis methods
Immunologic Techniques
Gene expression
Transcription Factors
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 14
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....7ce4fc7794b7e071cd1b6583b77b0929