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ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells
The DNA uptake of naturally competent bacteria has been attributed to the action of DNA uptake machineries resembling type IV pilus complexes. However, the protein(s) for pulling the DNA across the outer membrane of Gram-negative bacteria remain speculative. Here we show that the competence protein...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879209/ https://www.ncbi.nlm.nih.gov/pubmed/24391524 http://dx.doi.org/10.1371/journal.pgen.1004066 |
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author | Seitz, Patrick Pezeshgi Modarres, Hassan Borgeaud, Sandrine Bulushev, Roman D. Steinbock, Lorenz J. Radenovic, Aleksandra Dal Peraro, Matteo Blokesch, Melanie |
author_facet | Seitz, Patrick Pezeshgi Modarres, Hassan Borgeaud, Sandrine Bulushev, Roman D. Steinbock, Lorenz J. Radenovic, Aleksandra Dal Peraro, Matteo Blokesch, Melanie |
author_sort | Seitz, Patrick |
collection | PubMed |
description | The DNA uptake of naturally competent bacteria has been attributed to the action of DNA uptake machineries resembling type IV pilus complexes. However, the protein(s) for pulling the DNA across the outer membrane of Gram-negative bacteria remain speculative. Here we show that the competence protein ComEA binds incoming DNA in the periplasm of naturally competent Vibrio cholerae cells thereby promoting DNA uptake, possibly through ratcheting and entropic forces associated with ComEA binding. Using comparative modeling and molecular simulations, we projected the 3D structure and DNA-binding site of ComEA. These in silico predictions, combined with in vivo and in vitro validations of wild-type and site-directed modified variants of ComEA, suggested that ComEA is not solely a DNA receptor protein but plays a direct role in the DNA uptake process. Furthermore, we uncovered that ComEA homologs of other bacteria (both Gram-positive and Gram-negative) efficiently compensated for the absence of ComEA in V. cholerae, suggesting that the contribution of ComEA in the DNA uptake process might be conserved among naturally competent bacteria. |
format | Online Article Text |
id | pubmed-3879209 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-38792092014-01-03 ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells Seitz, Patrick Pezeshgi Modarres, Hassan Borgeaud, Sandrine Bulushev, Roman D. Steinbock, Lorenz J. Radenovic, Aleksandra Dal Peraro, Matteo Blokesch, Melanie PLoS Genet Research Article The DNA uptake of naturally competent bacteria has been attributed to the action of DNA uptake machineries resembling type IV pilus complexes. However, the protein(s) for pulling the DNA across the outer membrane of Gram-negative bacteria remain speculative. Here we show that the competence protein ComEA binds incoming DNA in the periplasm of naturally competent Vibrio cholerae cells thereby promoting DNA uptake, possibly through ratcheting and entropic forces associated with ComEA binding. Using comparative modeling and molecular simulations, we projected the 3D structure and DNA-binding site of ComEA. These in silico predictions, combined with in vivo and in vitro validations of wild-type and site-directed modified variants of ComEA, suggested that ComEA is not solely a DNA receptor protein but plays a direct role in the DNA uptake process. Furthermore, we uncovered that ComEA homologs of other bacteria (both Gram-positive and Gram-negative) efficiently compensated for the absence of ComEA in V. cholerae, suggesting that the contribution of ComEA in the DNA uptake process might be conserved among naturally competent bacteria. Public Library of Science 2014-01-02 /pmc/articles/PMC3879209/ /pubmed/24391524 http://dx.doi.org/10.1371/journal.pgen.1004066 Text en © 2014 Seitz et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Seitz, Patrick Pezeshgi Modarres, Hassan Borgeaud, Sandrine Bulushev, Roman D. Steinbock, Lorenz J. Radenovic, Aleksandra Dal Peraro, Matteo Blokesch, Melanie ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells |
title | ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells |
title_full | ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells |
title_fullStr | ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells |
title_full_unstemmed | ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells |
title_short | ComEA Is Essential for the Transfer of External DNA into the Periplasm in Naturally Transformable Vibrio cholerae Cells |
title_sort | comea is essential for the transfer of external dna into the periplasm in naturally transformable vibrio cholerae cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879209/ https://www.ncbi.nlm.nih.gov/pubmed/24391524 http://dx.doi.org/10.1371/journal.pgen.1004066 |
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