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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 (...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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American Society for Microbiology
2021
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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. |
format | Online Article Text |
id | pubmed-8546550 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
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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