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A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens

Periplasmic binding proteins (PBPs) in association with ABC transporters select and import a wide variety of ligands into bacterial cytoplasm. They can also take up toxic molecules, as observed in the case of the phytopathogen Agrobacterium tumefaciens strain C58. This organism contains a PBP called...

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Autores principales: El Sahili, Abbas, Li, Si-Zhe, Lang, Julien, Virus, Cornelia, Planamente, Sara, Ahmar, Mohammed, Guimaraes, Beatriz G., Aumont-Nicaise, Magali, Vigouroux, Armelle, Soulère, Laurent, Reader, John, Queneau, Yves, Faure, Denis, Moréra, Solange
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4526662/
https://www.ncbi.nlm.nih.gov/pubmed/26244338
http://dx.doi.org/10.1371/journal.ppat.1005071
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author El Sahili, Abbas
Li, Si-Zhe
Lang, Julien
Virus, Cornelia
Planamente, Sara
Ahmar, Mohammed
Guimaraes, Beatriz G.
Aumont-Nicaise, Magali
Vigouroux, Armelle
Soulère, Laurent
Reader, John
Queneau, Yves
Faure, Denis
Moréra, Solange
author_facet El Sahili, Abbas
Li, Si-Zhe
Lang, Julien
Virus, Cornelia
Planamente, Sara
Ahmar, Mohammed
Guimaraes, Beatriz G.
Aumont-Nicaise, Magali
Vigouroux, Armelle
Soulère, Laurent
Reader, John
Queneau, Yves
Faure, Denis
Moréra, Solange
author_sort El Sahili, Abbas
collection PubMed
description Periplasmic binding proteins (PBPs) in association with ABC transporters select and import a wide variety of ligands into bacterial cytoplasm. They can also take up toxic molecules, as observed in the case of the phytopathogen Agrobacterium tumefaciens strain C58. This organism contains a PBP called AccA that mediates the import of the antibiotic agrocin 84, as well as the opine agrocinopine A that acts as both a nutrient and a signalling molecule for the dissemination of virulence genes through quorum-sensing. Here, we characterized the binding mode of AccA using purified agrocin 84 and synthetic agrocinopine A by X-ray crystallography at very high resolution and performed affinity measurements. Structural and affinity analyses revealed that AccA recognizes an uncommon and specific motif, a pyranose-2-phosphate moiety which is present in both imported molecules via the L-arabinopyranose moiety in agrocinopine A and the D-glucopyranose moiety in agrocin 84. We hypothesized that AccA is a gateway allowing the import of any compound possessing a pyranose-2-phosphate motif at one end. This was structurally and functionally confirmed by experiments using four synthetic compounds: agrocinopine 3’-O-benzoate, L-arabinose-2-isopropylphosphate, L-arabinose-2-phosphate and D-glucose-2-phosphate. By combining affinity measurements and in vivo assays, we demonstrated that both L-arabinose-2-phosphate and D-glucose-2-phosphate, which are the AccF mediated degradation products of agrocinopine A and agrocin 84 respectively, interact with the master transcriptional regulator AccR and activate the quorum-sensing signal synthesis and Ti plasmid transfer in A. tumefaciens C58. Our findings shed light on the role of agrocinopine and antibiotic agrocin 84 on quorum-sensing regulation in A. tumefaciens and reveal how the PBP AccA acts as vehicle for the importation of both molecules by means of a key-recognition motif. It also opens future possibilities for the rational design of antibiotic and anti-virulence compounds against A. tumefaciens or other pathogens possessing similar PBPs.
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spelling pubmed-45266622015-08-12 A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens El Sahili, Abbas Li, Si-Zhe Lang, Julien Virus, Cornelia Planamente, Sara Ahmar, Mohammed Guimaraes, Beatriz G. Aumont-Nicaise, Magali Vigouroux, Armelle Soulère, Laurent Reader, John Queneau, Yves Faure, Denis Moréra, Solange PLoS Pathog Research Article Periplasmic binding proteins (PBPs) in association with ABC transporters select and import a wide variety of ligands into bacterial cytoplasm. They can also take up toxic molecules, as observed in the case of the phytopathogen Agrobacterium tumefaciens strain C58. This organism contains a PBP called AccA that mediates the import of the antibiotic agrocin 84, as well as the opine agrocinopine A that acts as both a nutrient and a signalling molecule for the dissemination of virulence genes through quorum-sensing. Here, we characterized the binding mode of AccA using purified agrocin 84 and synthetic agrocinopine A by X-ray crystallography at very high resolution and performed affinity measurements. Structural and affinity analyses revealed that AccA recognizes an uncommon and specific motif, a pyranose-2-phosphate moiety which is present in both imported molecules via the L-arabinopyranose moiety in agrocinopine A and the D-glucopyranose moiety in agrocin 84. We hypothesized that AccA is a gateway allowing the import of any compound possessing a pyranose-2-phosphate motif at one end. This was structurally and functionally confirmed by experiments using four synthetic compounds: agrocinopine 3’-O-benzoate, L-arabinose-2-isopropylphosphate, L-arabinose-2-phosphate and D-glucose-2-phosphate. By combining affinity measurements and in vivo assays, we demonstrated that both L-arabinose-2-phosphate and D-glucose-2-phosphate, which are the AccF mediated degradation products of agrocinopine A and agrocin 84 respectively, interact with the master transcriptional regulator AccR and activate the quorum-sensing signal synthesis and Ti plasmid transfer in A. tumefaciens C58. Our findings shed light on the role of agrocinopine and antibiotic agrocin 84 on quorum-sensing regulation in A. tumefaciens and reveal how the PBP AccA acts as vehicle for the importation of both molecules by means of a key-recognition motif. It also opens future possibilities for the rational design of antibiotic and anti-virulence compounds against A. tumefaciens or other pathogens possessing similar PBPs. Public Library of Science 2015-08-05 /pmc/articles/PMC4526662/ /pubmed/26244338 http://dx.doi.org/10.1371/journal.ppat.1005071 Text en © 2015 El Sahili 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
El Sahili, Abbas
Li, Si-Zhe
Lang, Julien
Virus, Cornelia
Planamente, Sara
Ahmar, Mohammed
Guimaraes, Beatriz G.
Aumont-Nicaise, Magali
Vigouroux, Armelle
Soulère, Laurent
Reader, John
Queneau, Yves
Faure, Denis
Moréra, Solange
A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens
title A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens
title_full A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens
title_fullStr A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens
title_full_unstemmed A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens
title_short A Pyranose-2-Phosphate Motif Is Responsible for Both Antibiotic Import and Quorum-Sensing Regulation in Agrobacterium tumefaciens
title_sort pyranose-2-phosphate motif is responsible for both antibiotic import and quorum-sensing regulation in agrobacterium tumefaciens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4526662/
https://www.ncbi.nlm.nih.gov/pubmed/26244338
http://dx.doi.org/10.1371/journal.ppat.1005071
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