Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris

The GntR family transcription regulator HpaR1 identified from Xanthomonas campestris pv. campestris has been previously shown to positively regulate the genes responsible for hypersensitive reaction and pathogenicity and to autorepress its own expression. Here, we demonstrated that HpaR1 is a global...

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Autores principales: Su, Hui-Zhao, Wu, Liu, Qi, Yan-Hua, Liu, Guo-Fang, Lu, Guang-Tao, Tang, Ji-Liang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4730234/
https://www.ncbi.nlm.nih.gov/pubmed/26818230
http://dx.doi.org/10.1038/srep19862
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author Su, Hui-Zhao
Wu, Liu
Qi, Yan-Hua
Liu, Guo-Fang
Lu, Guang-Tao
Tang, Ji-Liang
author_facet Su, Hui-Zhao
Wu, Liu
Qi, Yan-Hua
Liu, Guo-Fang
Lu, Guang-Tao
Tang, Ji-Liang
author_sort Su, Hui-Zhao
collection PubMed
description The GntR family transcription regulator HpaR1 identified from Xanthomonas campestris pv. campestris has been previously shown to positively regulate the genes responsible for hypersensitive reaction and pathogenicity and to autorepress its own expression. Here, we demonstrated that HpaR1 is a global regulator that positively regulates diverse biological processes, including xanthan polysaccharide production, extracellular enzyme activity, cell motility and tolerance to various stresses. To investigate the regulatory mechanisms of HpaR1, we began with xanthan polysaccharide production, which is governed by a cluster of gum genes. These are directed by the gumB promoter. Disruption of HpaR1 significantly reduced gumB transcription and an electrophoretic mobility shift assay demonstrated that HpaR1 interacts directly with gumB promoter. DNase I footprint analysis revealed that HpaR1 and RNA polymerase were bound to the sequences extending from −21 to +10 and −41 to +29 relative to the transcription initiation site of gumB, respectively. Furthermore, in vitro transcription assays showed that HpaR1 facilitated the binding of RNA polymerase to gumB promoter, leading to an enhancement of its transcription. These results suggest that HpaR1 regulates gumB transcription via a mechanism similar but different to what was found, until now, to only be used by some MerR family transcription activators.
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spelling pubmed-47302342016-02-03 Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris Su, Hui-Zhao Wu, Liu Qi, Yan-Hua Liu, Guo-Fang Lu, Guang-Tao Tang, Ji-Liang Sci Rep Article The GntR family transcription regulator HpaR1 identified from Xanthomonas campestris pv. campestris has been previously shown to positively regulate the genes responsible for hypersensitive reaction and pathogenicity and to autorepress its own expression. Here, we demonstrated that HpaR1 is a global regulator that positively regulates diverse biological processes, including xanthan polysaccharide production, extracellular enzyme activity, cell motility and tolerance to various stresses. To investigate the regulatory mechanisms of HpaR1, we began with xanthan polysaccharide production, which is governed by a cluster of gum genes. These are directed by the gumB promoter. Disruption of HpaR1 significantly reduced gumB transcription and an electrophoretic mobility shift assay demonstrated that HpaR1 interacts directly with gumB promoter. DNase I footprint analysis revealed that HpaR1 and RNA polymerase were bound to the sequences extending from −21 to +10 and −41 to +29 relative to the transcription initiation site of gumB, respectively. Furthermore, in vitro transcription assays showed that HpaR1 facilitated the binding of RNA polymerase to gumB promoter, leading to an enhancement of its transcription. These results suggest that HpaR1 regulates gumB transcription via a mechanism similar but different to what was found, until now, to only be used by some MerR family transcription activators. Nature Publishing Group 2016-01-28 /pmc/articles/PMC4730234/ /pubmed/26818230 http://dx.doi.org/10.1038/srep19862 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Su, Hui-Zhao
Wu, Liu
Qi, Yan-Hua
Liu, Guo-Fang
Lu, Guang-Tao
Tang, Ji-Liang
Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris
title Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris
title_full Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris
title_fullStr Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris
title_full_unstemmed Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris
title_short Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris
title_sort characterization of the gntr family regulator hpar1 of the crucifer black rot pathogen xanthomonas campestris pathovar campestris
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4730234/
https://www.ncbi.nlm.nih.gov/pubmed/26818230
http://dx.doi.org/10.1038/srep19862
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