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Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR

Cholera is a diarrheal disease caused by the Gram-negative bacterium Vibrio cholerae. To reach the surface of intestinal epithelial cells, proliferate, and cause disease, V. cholerae tightly regulates the production of virulence factors such as cholera toxin (ctxAB) and the toxin-coregulated pilus (...

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Autores principales: Calkins, A. L., Demey, L. M., Karslake, J. D., Donarski, E. D., Biteen, J. S., DiRita, V. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8546550/
https://www.ncbi.nlm.nih.gov/pubmed/34488449
http://dx.doi.org/10.1128/mBio.02213-21
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author Calkins, A. L.
Demey, L. M.
Karslake, J. D.
Donarski, E. D.
Biteen, J. S.
DiRita, V. J.
author_facet Calkins, A. L.
Demey, L. M.
Karslake, J. D.
Donarski, E. D.
Biteen, J. S.
DiRita, V. J.
author_sort Calkins, A. L.
collection PubMed
description Cholera is a diarrheal disease caused by the Gram-negative bacterium Vibrio cholerae. To reach the surface of intestinal epithelial cells, proliferate, and cause disease, V. cholerae tightly regulates the production of virulence factors such as cholera toxin (ctxAB) and the toxin-coregulated pilus (tcpA-F). ToxT is directly responsible for regulating these major virulence factors while TcpP and ToxR indirectly regulate virulence factor production by stimulating toxT expression. TcpP and ToxR are membrane-localized transcription activators (MLTAs) required to activate toxT expression. To gain a deeper understanding of how MLTAs identify promoter DNA while in the membrane, we tracked the dynamics of single TcpP-PAmCherry molecules in live cells using photoactivated localization microscopy and identified heterogeneous diffusion patterns. Our results provide evidence that (i) TcpP exists in three biophysical states (fast diffusion, intermediate diffusion, and slow diffusion), (ii) TcpP transitions between these different diffusion states, (iii) TcpP molecules in the slow diffusion state are interacting with the toxT promoter, and (iv) ToxR is not essential for TcpP to localize the toxT promoter. These data refine the current model of cooperativity between TcpP and ToxR in stimulating toxT expression and demonstrate that TcpP locates the toxT promoter independently of ToxR.
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spelling pubmed-85465502021-11-04 Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR Calkins, A. L. Demey, L. M. Karslake, J. D. Donarski, E. D. Biteen, J. S. DiRita, V. J. mBio Research Article Cholera is a diarrheal disease caused by the Gram-negative bacterium Vibrio cholerae. To reach the surface of intestinal epithelial cells, proliferate, and cause disease, V. cholerae tightly regulates the production of virulence factors such as cholera toxin (ctxAB) and the toxin-coregulated pilus (tcpA-F). ToxT is directly responsible for regulating these major virulence factors while TcpP and ToxR indirectly regulate virulence factor production by stimulating toxT expression. TcpP and ToxR are membrane-localized transcription activators (MLTAs) required to activate toxT expression. To gain a deeper understanding of how MLTAs identify promoter DNA while in the membrane, we tracked the dynamics of single TcpP-PAmCherry molecules in live cells using photoactivated localization microscopy and identified heterogeneous diffusion patterns. Our results provide evidence that (i) TcpP exists in three biophysical states (fast diffusion, intermediate diffusion, and slow diffusion), (ii) TcpP transitions between these different diffusion states, (iii) TcpP molecules in the slow diffusion state are interacting with the toxT promoter, and (iv) ToxR is not essential for TcpP to localize the toxT promoter. These data refine the current model of cooperativity between TcpP and ToxR in stimulating toxT expression and demonstrate that TcpP locates the toxT promoter independently of ToxR. American Society for Microbiology 2021-09-07 /pmc/articles/PMC8546550/ /pubmed/34488449 http://dx.doi.org/10.1128/mBio.02213-21 Text en Copyright © 2021 Calkins et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Calkins, A. L.
Demey, L. M.
Karslake, J. D.
Donarski, E. D.
Biteen, J. S.
DiRita, V. J.
Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR
title Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR
title_full Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR
title_fullStr Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR
title_full_unstemmed Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR
title_short Independent Promoter Recognition by TcpP Precedes Cooperative Promoter Activation by TcpP and ToxR
title_sort independent promoter recognition by tcpp precedes cooperative promoter activation by tcpp and toxr
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8546550/
https://www.ncbi.nlm.nih.gov/pubmed/34488449
http://dx.doi.org/10.1128/mBio.02213-21
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