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The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus

Dichelobacter nodosus is the principal causative agent of ovine footrot and its extracellular proteases are major virulence factors. Virulent isolates of D. nodosus secrete three subtilisin-like serine proteases: AprV2, AprV5 and BprV. These enzymes are each synthesized as precursor molecules that i...

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Autores principales: Han, Xiaoyan, Kennan, Ruth M., Steer, David L., Smith, A. Ian, Whisstock, James C., Rood, Julian I.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3480457/
https://www.ncbi.nlm.nih.gov/pubmed/23112874
http://dx.doi.org/10.1371/journal.pone.0047932
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author Han, Xiaoyan
Kennan, Ruth M.
Steer, David L.
Smith, A. Ian
Whisstock, James C.
Rood, Julian I.
author_facet Han, Xiaoyan
Kennan, Ruth M.
Steer, David L.
Smith, A. Ian
Whisstock, James C.
Rood, Julian I.
author_sort Han, Xiaoyan
collection PubMed
description Dichelobacter nodosus is the principal causative agent of ovine footrot and its extracellular proteases are major virulence factors. Virulent isolates of D. nodosus secrete three subtilisin-like serine proteases: AprV2, AprV5 and BprV. These enzymes are each synthesized as precursor molecules that include a signal (pre-) peptide, a pro-peptide and a C-terminal extension, which are processed to produce the mature active forms. The function of the C-terminal regions of these proteases and the mechanism of protease processing and secretion are unknown. AprV5 contributes to most of the protease activity secreted by D. nodosus. To understand the role of the C-terminal extension of AprV5, we constructed a series of C-terminal-deletion mutants in D. nodosus by allelic exchange. The proteases present in the resultant mutants and their complemented derivatives were examined by protease zymogram analysis, western blotting and mass spectrometry. The results showed that the C-terminal region of AprV5 is required for the normal expression of protease activity, deletion of this region led to a delay in the processing of these enzymes. D. nodosus is an unusual bacterium in that it produces three closely related extracellular serine proteases. We have now shown that one of these enzymes, AprV5, is responsible for its own maturation, and for the optimal cleavage of AprV2 and BprV, to their mature active forms. These studies have increased our understanding of how this important pathogen processes these virulence-associated extracellular proteases and secretes them into its external environment.
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spelling pubmed-34804572012-10-30 The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus Han, Xiaoyan Kennan, Ruth M. Steer, David L. Smith, A. Ian Whisstock, James C. Rood, Julian I. PLoS One Research Article Dichelobacter nodosus is the principal causative agent of ovine footrot and its extracellular proteases are major virulence factors. Virulent isolates of D. nodosus secrete three subtilisin-like serine proteases: AprV2, AprV5 and BprV. These enzymes are each synthesized as precursor molecules that include a signal (pre-) peptide, a pro-peptide and a C-terminal extension, which are processed to produce the mature active forms. The function of the C-terminal regions of these proteases and the mechanism of protease processing and secretion are unknown. AprV5 contributes to most of the protease activity secreted by D. nodosus. To understand the role of the C-terminal extension of AprV5, we constructed a series of C-terminal-deletion mutants in D. nodosus by allelic exchange. The proteases present in the resultant mutants and their complemented derivatives were examined by protease zymogram analysis, western blotting and mass spectrometry. The results showed that the C-terminal region of AprV5 is required for the normal expression of protease activity, deletion of this region led to a delay in the processing of these enzymes. D. nodosus is an unusual bacterium in that it produces three closely related extracellular serine proteases. We have now shown that one of these enzymes, AprV5, is responsible for its own maturation, and for the optimal cleavage of AprV2 and BprV, to their mature active forms. These studies have increased our understanding of how this important pathogen processes these virulence-associated extracellular proteases and secretes them into its external environment. Public Library of Science 2012-10-24 /pmc/articles/PMC3480457/ /pubmed/23112874 http://dx.doi.org/10.1371/journal.pone.0047932 Text en © 2012 Han 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
Han, Xiaoyan
Kennan, Ruth M.
Steer, David L.
Smith, A. Ian
Whisstock, James C.
Rood, Julian I.
The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus
title The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus
title_full The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus
title_fullStr The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus
title_full_unstemmed The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus
title_short The AprV5 Subtilase Is Required for the Optimal Processing of All Three Extracellular Serine Proteases from Dichelobacter nodosus
title_sort aprv5 subtilase is required for the optimal processing of all three extracellular serine proteases from dichelobacter nodosus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3480457/
https://www.ncbi.nlm.nih.gov/pubmed/23112874
http://dx.doi.org/10.1371/journal.pone.0047932
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