The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.

A transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3',5'-cyclic diguanylic acid (c-di-GMP) and encoded a calcium-binding protein CabA...

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Main Authors: Jin Hwan Park, Youmi Jo, Song Yee Jang, Haenaem Kwon, Yasuhiko Irie, Matthew R Parsek, Myung Hee Kim, Sang Ho Choi
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2015-09-01
Series:PLoS Pathogens
Online Access:http://europepmc.org/articles/PMC4584020?pdf=render
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author Jin Hwan Park
Youmi Jo
Song Yee Jang
Haenaem Kwon
Yasuhiko Irie
Matthew R Parsek
Myung Hee Kim
Sang Ho Choi
author_facet Jin Hwan Park
Youmi Jo
Song Yee Jang
Haenaem Kwon
Yasuhiko Irie
Matthew R Parsek
Myung Hee Kim
Sang Ho Choi
author_sort Jin Hwan Park
collection DOAJ
description A transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3',5'-cyclic diguanylic acid (c-di-GMP) and encoded a calcium-binding protein CabA. Comparison of the biofilms produced by the cabA mutant and its parent strain JN111 in microtiter plates using crystal-violet staining demonstrated that CabA contributed to biofilm formation in a calcium-dependent manner under elevated c-di-GMP conditions. Genetic and biochemical analyses revealed that CabA was secreted to the cell exterior through functional CabB and CabC, distributed throughout the biofilm matrix, and produced as the biofilm matured. These results, together with the observation that CabA also contributes to the development of rugose colony morphology, indicated that CabA is a matrix-associated protein required for maturation, rather than adhesion involved in the initial attachment, of biofilms. Microscopic comparison of the structure of biofilms produced by JN111 and the cabA mutant demonstrated that CabA is an extracellular matrix component essential for the development of the mature biofilm structures in flow cells and on oyster shells. Exogenously providing purified CabA restored the biofilm- and rugose colony-forming abilities of the cabA mutant when calcium was available. Circular dichroism and size exclusion analyses revealed that calcium binding induces CabA conformational changes which may lead to multimerization. Extracellular complementation experiments revealed that CabA can assemble a functional matrix only when exopolysaccharides coexist. Consequently, the combined results suggested that CabA is a structural protein of the extracellular matrix and multimerizes to a conformation functional in building robust biofilms, which may render V. vulnificus to survive in hostile environments and reach a concentrated infective dose.
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spelling doaj.art-20231f8c712b4946af861f38f8339a092022-12-22T03:42:28ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742015-09-01119e100519210.1371/journal.ppat.1005192The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.Jin Hwan ParkYoumi JoSong Yee JangHaenaem KwonYasuhiko IrieMatthew R ParsekMyung Hee KimSang Ho ChoiA transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3',5'-cyclic diguanylic acid (c-di-GMP) and encoded a calcium-binding protein CabA. Comparison of the biofilms produced by the cabA mutant and its parent strain JN111 in microtiter plates using crystal-violet staining demonstrated that CabA contributed to biofilm formation in a calcium-dependent manner under elevated c-di-GMP conditions. Genetic and biochemical analyses revealed that CabA was secreted to the cell exterior through functional CabB and CabC, distributed throughout the biofilm matrix, and produced as the biofilm matured. These results, together with the observation that CabA also contributes to the development of rugose colony morphology, indicated that CabA is a matrix-associated protein required for maturation, rather than adhesion involved in the initial attachment, of biofilms. Microscopic comparison of the structure of biofilms produced by JN111 and the cabA mutant demonstrated that CabA is an extracellular matrix component essential for the development of the mature biofilm structures in flow cells and on oyster shells. Exogenously providing purified CabA restored the biofilm- and rugose colony-forming abilities of the cabA mutant when calcium was available. Circular dichroism and size exclusion analyses revealed that calcium binding induces CabA conformational changes which may lead to multimerization. Extracellular complementation experiments revealed that CabA can assemble a functional matrix only when exopolysaccharides coexist. Consequently, the combined results suggested that CabA is a structural protein of the extracellular matrix and multimerizes to a conformation functional in building robust biofilms, which may render V. vulnificus to survive in hostile environments and reach a concentrated infective dose.http://europepmc.org/articles/PMC4584020?pdf=render
spellingShingle Jin Hwan Park
Youmi Jo
Song Yee Jang
Haenaem Kwon
Yasuhiko Irie
Matthew R Parsek
Myung Hee Kim
Sang Ho Choi
The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.
PLoS Pathogens
title The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.
title_full The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.
title_fullStr The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.
title_full_unstemmed The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.
title_short The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.
title_sort cababc operon essential for biofilm and rugose colony development in vibrio vulnificus
url http://europepmc.org/articles/PMC4584020?pdf=render
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