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Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007

BACKGROUND: Ruminococcus flavefaciens is an important fibre-degrading bacterium found in the mammalian gut. Cellulolytic strains from the bovine rumen have been shown to produce complex cellulosome structures that are associated with the cell surface. R. flavefaciens 007 is a highly cellulolytic str...

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Autores principales: Vodovnik, Maša, Duncan, Sylvia H., Reid, Martin D., Cantlay, Louise, Turner, Keith, Parkhill, Julian, Lamed, Raphael, Yeoman, Carl J., Miller, Margret E. Berg., White, Bryan A., Bayer, Edward A., Marinšek-Logar, Romana, Flint, Harry J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3672088/
https://www.ncbi.nlm.nih.gov/pubmed/23750253
http://dx.doi.org/10.1371/journal.pone.0065333
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author Vodovnik, Maša
Duncan, Sylvia H.
Reid, Martin D.
Cantlay, Louise
Turner, Keith
Parkhill, Julian
Lamed, Raphael
Yeoman, Carl J.
Miller, Margret E. Berg.
White, Bryan A.
Bayer, Edward A.
Marinšek-Logar, Romana
Flint, Harry J.
author_facet Vodovnik, Maša
Duncan, Sylvia H.
Reid, Martin D.
Cantlay, Louise
Turner, Keith
Parkhill, Julian
Lamed, Raphael
Yeoman, Carl J.
Miller, Margret E. Berg.
White, Bryan A.
Bayer, Edward A.
Marinšek-Logar, Romana
Flint, Harry J.
author_sort Vodovnik, Maša
collection PubMed
description BACKGROUND: Ruminococcus flavefaciens is an important fibre-degrading bacterium found in the mammalian gut. Cellulolytic strains from the bovine rumen have been shown to produce complex cellulosome structures that are associated with the cell surface. R. flavefaciens 007 is a highly cellulolytic strain whose ability to degrade dewaxed cotton, but not Avicel cellulose, was lost following initial isolation in the variant 007S. The ability was recovered after serial subculture to give the cotton-degrading strain 007C. This has allowed us to investigate the factors required for degradation of this particularly recalcitrant form of cellulose. METHODOLOGY/PRINCIPAL FINDINGS: The major proteins associated with the bacterial cell surface and with the culture supernatant were analyzed for R. flavefaciens 007S and 007C grown with cellobiose, xylan or Avicel cellulose as energy sources. Identification of the proteins was enabled by a draft genome sequence obtained for 007C. Among supernatant proteins a cellulosomal GH48 hydrolase, a rubrerthyrin-like protein and a protein with type IV pili N-terminal domain were the most strongly up-regulated in 007C cultures grown on Avicel compared with cellobiose. Strain 007S also showed substrate-related changes, but supernatant expression of the Pil protein and rubrerythrin in particular were markedly lower in 007S than in 007C during growth on Avicel. CONCLUSIONS/SIGNIFICANCE: This study provides new information on the extracellular proteome of R. flavefaciens and its regulation in response to different growth substrates. Furthermore it suggests that the cotton cellulose non-degrading strain (007S) has altered regulation of multiple proteins that may be required for breakdown of cotton cellulose. One of these, the type IV pilus was previously shown to play a role in adhesion to cellulose in R. albus, and a related pilin protein was identified here for the first time as a major extracellular protein in R. flavefaciens.
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spelling pubmed-36720882013-06-07 Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007 Vodovnik, Maša Duncan, Sylvia H. Reid, Martin D. Cantlay, Louise Turner, Keith Parkhill, Julian Lamed, Raphael Yeoman, Carl J. Miller, Margret E. Berg. White, Bryan A. Bayer, Edward A. Marinšek-Logar, Romana Flint, Harry J. PLoS One Research Article BACKGROUND: Ruminococcus flavefaciens is an important fibre-degrading bacterium found in the mammalian gut. Cellulolytic strains from the bovine rumen have been shown to produce complex cellulosome structures that are associated with the cell surface. R. flavefaciens 007 is a highly cellulolytic strain whose ability to degrade dewaxed cotton, but not Avicel cellulose, was lost following initial isolation in the variant 007S. The ability was recovered after serial subculture to give the cotton-degrading strain 007C. This has allowed us to investigate the factors required for degradation of this particularly recalcitrant form of cellulose. METHODOLOGY/PRINCIPAL FINDINGS: The major proteins associated with the bacterial cell surface and with the culture supernatant were analyzed for R. flavefaciens 007S and 007C grown with cellobiose, xylan or Avicel cellulose as energy sources. Identification of the proteins was enabled by a draft genome sequence obtained for 007C. Among supernatant proteins a cellulosomal GH48 hydrolase, a rubrerthyrin-like protein and a protein with type IV pili N-terminal domain were the most strongly up-regulated in 007C cultures grown on Avicel compared with cellobiose. Strain 007S also showed substrate-related changes, but supernatant expression of the Pil protein and rubrerythrin in particular were markedly lower in 007S than in 007C during growth on Avicel. CONCLUSIONS/SIGNIFICANCE: This study provides new information on the extracellular proteome of R. flavefaciens and its regulation in response to different growth substrates. Furthermore it suggests that the cotton cellulose non-degrading strain (007S) has altered regulation of multiple proteins that may be required for breakdown of cotton cellulose. One of these, the type IV pilus was previously shown to play a role in adhesion to cellulose in R. albus, and a related pilin protein was identified here for the first time as a major extracellular protein in R. flavefaciens. Public Library of Science 2013-06-04 /pmc/articles/PMC3672088/ /pubmed/23750253 http://dx.doi.org/10.1371/journal.pone.0065333 Text en © 2013 Vodovnik 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
Vodovnik, Maša
Duncan, Sylvia H.
Reid, Martin D.
Cantlay, Louise
Turner, Keith
Parkhill, Julian
Lamed, Raphael
Yeoman, Carl J.
Miller, Margret E. Berg.
White, Bryan A.
Bayer, Edward A.
Marinšek-Logar, Romana
Flint, Harry J.
Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007
title Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007
title_full Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007
title_fullStr Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007
title_full_unstemmed Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007
title_short Expression of Cellulosome Components and Type IV Pili within the Extracellular Proteome of Ruminococcus flavefaciens 007
title_sort expression of cellulosome components and type iv pili within the extracellular proteome of ruminococcus flavefaciens 007
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3672088/
https://www.ncbi.nlm.nih.gov/pubmed/23750253
http://dx.doi.org/10.1371/journal.pone.0065333
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