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Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF

The biofilm matrix of the rhizospheric bacterium Pseudomonas putida consists mainly of a proteinaceous component. The two largest P. putida proteins, adhesins LapA and LapF, are involved in biofilm development but prevail in different developmental stages of the biofilm matrix. LapA is abundant in t...

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Autores principales: Lahesaare, Andrio, Moor, Hanna, Kivisaar, Maia, Teras, Riho
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4278767/
https://www.ncbi.nlm.nih.gov/pubmed/25545773
http://dx.doi.org/10.1371/journal.pone.0115901
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author Lahesaare, Andrio
Moor, Hanna
Kivisaar, Maia
Teras, Riho
author_facet Lahesaare, Andrio
Moor, Hanna
Kivisaar, Maia
Teras, Riho
author_sort Lahesaare, Andrio
collection PubMed
description The biofilm matrix of the rhizospheric bacterium Pseudomonas putida consists mainly of a proteinaceous component. The two largest P. putida proteins, adhesins LapA and LapF, are involved in biofilm development but prevail in different developmental stages of the biofilm matrix. LapA is abundant in the initial stage of biofilm formation whereas LapF is found in the mature biofilm. Although the transcriptional regulation of the adhesins is not exhaustively studied, some factors that can be involved in their regulation have been described. For example, RpoS, the major stress response sigma factor, activates, and Fis represses LapF expression. This study focused on the LapF expression control by Fis. Indeed, using DNase I footprint analysis a Fis binding site Fis-F2 was located 150 bp upstream of the lapF gene coding sequence. The mapped 5′ end of the lapF mRNA localized the promoter to the same region, overlapping with the Fis binding site Fis-F2. Monitoring the lapF promoter activity by a β-galactosidase assay revealed that Fis overexpression causes a 4-fold decrease in the transcriptional activity. Furthermore, mutations that diminished Fis binding to the Fis-F2 site abolished the repression of the lapF promoter. Thus, these data suggest that Fis is involved in the biofilm regulation via repression of LapF expression.
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spelling pubmed-42787672015-01-05 Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF Lahesaare, Andrio Moor, Hanna Kivisaar, Maia Teras, Riho PLoS One Research Article The biofilm matrix of the rhizospheric bacterium Pseudomonas putida consists mainly of a proteinaceous component. The two largest P. putida proteins, adhesins LapA and LapF, are involved in biofilm development but prevail in different developmental stages of the biofilm matrix. LapA is abundant in the initial stage of biofilm formation whereas LapF is found in the mature biofilm. Although the transcriptional regulation of the adhesins is not exhaustively studied, some factors that can be involved in their regulation have been described. For example, RpoS, the major stress response sigma factor, activates, and Fis represses LapF expression. This study focused on the LapF expression control by Fis. Indeed, using DNase I footprint analysis a Fis binding site Fis-F2 was located 150 bp upstream of the lapF gene coding sequence. The mapped 5′ end of the lapF mRNA localized the promoter to the same region, overlapping with the Fis binding site Fis-F2. Monitoring the lapF promoter activity by a β-galactosidase assay revealed that Fis overexpression causes a 4-fold decrease in the transcriptional activity. Furthermore, mutations that diminished Fis binding to the Fis-F2 site abolished the repression of the lapF promoter. Thus, these data suggest that Fis is involved in the biofilm regulation via repression of LapF expression. Public Library of Science 2014-12-29 /pmc/articles/PMC4278767/ /pubmed/25545773 http://dx.doi.org/10.1371/journal.pone.0115901 Text en © 2014 Lahesaare 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
Lahesaare, Andrio
Moor, Hanna
Kivisaar, Maia
Teras, Riho
Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF
title Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF
title_full Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF
title_fullStr Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF
title_full_unstemmed Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF
title_short Pseudomonas putida Fis Binds to the lapF Promoter In Vitro and Represses the Expression of LapF
title_sort pseudomonas putida fis binds to the lapf promoter in vitro and represses the expression of lapf
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4278767/
https://www.ncbi.nlm.nih.gov/pubmed/25545773
http://dx.doi.org/10.1371/journal.pone.0115901
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