Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens

Shin-You Ding, Marco T Rincon, Raphael Lamed, Jennifer C Martin, Sheila I. McCrae, Vincenzo Aurilia, Yuval Shoham, Edward A. Bayer, Harry J. Flint

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

Two tandem cellulosome-associated genes were identified in the cellulolytic rumen bacterium, Ruminococcus flavefaciens. The deduced gene products represent multimodular scaffoldin-related proteins (termed ScaA and ScaB), both of which include several copies of explicit cellulosome signature sequences. The scaB gene was completely sequenced, and its upstream neighbor scaA was partially sequenced. The sequenced portion of scaA contains repeating cohesin modules and a C-terminal dockerin domain. ScaB contains seven relatively divergent cohesin modules, two extremely long T-rich linkers, and a C-terminal domain of unknown function. Collectively, the cohesins of ScaA and ScaB are phylogenetically distinct from the previously described type I and type II cohesins, and we propose that they define a new group, which we designated here type III cohesins, Selected modules from both genes were overexpressed in Escherichia coli, and the recombinant proteins were used as probes in affinity-blotting experiments. The results strongly indicate that ScaA serves as a cellulosomal scaffoldin-like protein for several R. flavefaciens enzymes. The data are supported by the direct interaction of a recombinant ScaA cohesin with an expressed dockerin-containing enzyme construct from the same bacterium. The evidence also demonstrates that the ScaA dockerin binds to a specialized cohesin(s) on ScaB, suggesting that ScaB may act as an anchoring protein, linked either directly or indirectly to the bacterial cell surface. This study is the first direct demonstration in a cellulolytic rumen bacterium of a cellulosome system, mediated by distinctive cohesin-dockerin interactions.

Original languageEnglish
Pages (from-to)1945-1953
Number of pages9
JournalJournal of Bacteriology
Volume183
Issue number6
DOIs
Publication statusPublished - Mar 2001

Keywords

  • clostridium-thermocellum
  • binding-protein
  • subcellular-localization
  • secondary structure
  • sequence-analysis
  • dockerin domain
  • neural-networks
  • cell-surface
  • rumen
  • albus

Cite this

Ding, S-Y., Rincon, M. T., Lamed, R., Martin, J. C., McCrae, S. I., Aurilia, V., ... Flint, H. J. (2001). Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens. Journal of Bacteriology, 183(6), 1945-1953. https://doi.org/10.1128/JB.183.6.1945-1953.2001

Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens. / Ding, Shin-You; Rincon, Marco T ; Lamed, Raphael; Martin, Jennifer C; McCrae, Sheila I.; Aurilia, Vincenzo; Shoham, Yuval; Bayer, Edward A.; Flint, Harry J.

In: Journal of Bacteriology, Vol. 183, No. 6, 03.2001, p. 1945-1953.

Research output: Contribution to journalArticle

Ding, S-Y, Rincon, MT, Lamed, R, Martin, JC, McCrae, SI, Aurilia, V, Shoham, Y, Bayer, EA & Flint, HJ 2001, 'Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens', Journal of Bacteriology, vol. 183, no. 6, pp. 1945-1953. https://doi.org/10.1128/JB.183.6.1945-1953.2001
Ding S-Y, Rincon MT, Lamed R, Martin JC, McCrae SI, Aurilia V et al. Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens. Journal of Bacteriology. 2001 Mar;183(6):1945-1953. https://doi.org/10.1128/JB.183.6.1945-1953.2001
Ding, Shin-You ; Rincon, Marco T ; Lamed, Raphael ; Martin, Jennifer C ; McCrae, Sheila I. ; Aurilia, Vincenzo ; Shoham, Yuval ; Bayer, Edward A. ; Flint, Harry J. / Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens. In: Journal of Bacteriology. 2001 ; Vol. 183, No. 6. pp. 1945-1953.
@article{b88d72bf57c74f9c8e686ff94af8807c,
title = "Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens",
abstract = "Two tandem cellulosome-associated genes were identified in the cellulolytic rumen bacterium, Ruminococcus flavefaciens. The deduced gene products represent multimodular scaffoldin-related proteins (termed ScaA and ScaB), both of which include several copies of explicit cellulosome signature sequences. The scaB gene was completely sequenced, and its upstream neighbor scaA was partially sequenced. The sequenced portion of scaA contains repeating cohesin modules and a C-terminal dockerin domain. ScaB contains seven relatively divergent cohesin modules, two extremely long T-rich linkers, and a C-terminal domain of unknown function. Collectively, the cohesins of ScaA and ScaB are phylogenetically distinct from the previously described type I and type II cohesins, and we propose that they define a new group, which we designated here type III cohesins, Selected modules from both genes were overexpressed in Escherichia coli, and the recombinant proteins were used as probes in affinity-blotting experiments. The results strongly indicate that ScaA serves as a cellulosomal scaffoldin-like protein for several R. flavefaciens enzymes. The data are supported by the direct interaction of a recombinant ScaA cohesin with an expressed dockerin-containing enzyme construct from the same bacterium. The evidence also demonstrates that the ScaA dockerin binds to a specialized cohesin(s) on ScaB, suggesting that ScaB may act as an anchoring protein, linked either directly or indirectly to the bacterial cell surface. This study is the first direct demonstration in a cellulolytic rumen bacterium of a cellulosome system, mediated by distinctive cohesin-dockerin interactions.",
keywords = "clostridium-thermocellum, binding-protein, subcellular-localization, secondary structure, sequence-analysis, dockerin domain, neural-networks, cell-surface, rumen, albus",
author = "Shin-You Ding and Rincon, {Marco T} and Raphael Lamed and Martin, {Jennifer C} and McCrae, {Sheila I.} and Vincenzo Aurilia and Yuval Shoham and Bayer, {Edward A.} and Flint, {Harry J.}",
year = "2001",
month = "3",
doi = "10.1128/JB.183.6.1945-1953.2001",
language = "English",
volume = "183",
pages = "1945--1953",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "6",

}

TY - JOUR

T1 - Cellulosomal scaffoldin-like proteins from Ruminococcus flavefaciens

AU - Ding, Shin-You

AU - Rincon, Marco T

AU - Lamed, Raphael

AU - Martin, Jennifer C

AU - McCrae, Sheila I.

AU - Aurilia, Vincenzo

AU - Shoham, Yuval

AU - Bayer, Edward A.

AU - Flint, Harry J.

PY - 2001/3

Y1 - 2001/3

N2 - Two tandem cellulosome-associated genes were identified in the cellulolytic rumen bacterium, Ruminococcus flavefaciens. The deduced gene products represent multimodular scaffoldin-related proteins (termed ScaA and ScaB), both of which include several copies of explicit cellulosome signature sequences. The scaB gene was completely sequenced, and its upstream neighbor scaA was partially sequenced. The sequenced portion of scaA contains repeating cohesin modules and a C-terminal dockerin domain. ScaB contains seven relatively divergent cohesin modules, two extremely long T-rich linkers, and a C-terminal domain of unknown function. Collectively, the cohesins of ScaA and ScaB are phylogenetically distinct from the previously described type I and type II cohesins, and we propose that they define a new group, which we designated here type III cohesins, Selected modules from both genes were overexpressed in Escherichia coli, and the recombinant proteins were used as probes in affinity-blotting experiments. The results strongly indicate that ScaA serves as a cellulosomal scaffoldin-like protein for several R. flavefaciens enzymes. The data are supported by the direct interaction of a recombinant ScaA cohesin with an expressed dockerin-containing enzyme construct from the same bacterium. The evidence also demonstrates that the ScaA dockerin binds to a specialized cohesin(s) on ScaB, suggesting that ScaB may act as an anchoring protein, linked either directly or indirectly to the bacterial cell surface. This study is the first direct demonstration in a cellulolytic rumen bacterium of a cellulosome system, mediated by distinctive cohesin-dockerin interactions.

AB - Two tandem cellulosome-associated genes were identified in the cellulolytic rumen bacterium, Ruminococcus flavefaciens. The deduced gene products represent multimodular scaffoldin-related proteins (termed ScaA and ScaB), both of which include several copies of explicit cellulosome signature sequences. The scaB gene was completely sequenced, and its upstream neighbor scaA was partially sequenced. The sequenced portion of scaA contains repeating cohesin modules and a C-terminal dockerin domain. ScaB contains seven relatively divergent cohesin modules, two extremely long T-rich linkers, and a C-terminal domain of unknown function. Collectively, the cohesins of ScaA and ScaB are phylogenetically distinct from the previously described type I and type II cohesins, and we propose that they define a new group, which we designated here type III cohesins, Selected modules from both genes were overexpressed in Escherichia coli, and the recombinant proteins were used as probes in affinity-blotting experiments. The results strongly indicate that ScaA serves as a cellulosomal scaffoldin-like protein for several R. flavefaciens enzymes. The data are supported by the direct interaction of a recombinant ScaA cohesin with an expressed dockerin-containing enzyme construct from the same bacterium. The evidence also demonstrates that the ScaA dockerin binds to a specialized cohesin(s) on ScaB, suggesting that ScaB may act as an anchoring protein, linked either directly or indirectly to the bacterial cell surface. This study is the first direct demonstration in a cellulolytic rumen bacterium of a cellulosome system, mediated by distinctive cohesin-dockerin interactions.

KW - clostridium-thermocellum

KW - binding-protein

KW - subcellular-localization

KW - secondary structure

KW - sequence-analysis

KW - dockerin domain

KW - neural-networks

KW - cell-surface

KW - rumen

KW - albus

U2 - 10.1128/JB.183.6.1945-1953.2001

DO - 10.1128/JB.183.6.1945-1953.2001

M3 - Article

VL - 183

SP - 1945

EP - 1953

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 6

ER -