A Tau Class Glutathione-S-Transferase is Involved in Trans-Resveratrol Transport Out of Grapevine Cells

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Title: A Tau Class Glutathione-S-Transferase is Involved in Trans-Resveratrol Transport Out of Grapevine Cells
Authors: Martínez Márquez, Ascensión | Martínez Esteso, María José | Vilella Antón, María Teresa | Sellés Marchart, Susana | Morante Carriel, Jaime | Hurtado-Gaitán, Elías | Palazon, Javier | Bru-Martinez, Roque
Research Group/s: Proteómica y Genómica Funcional de Plantas
Center, Department or Service: Universidad de Alicante. Departamento de Agroquímica y Bioquímica
Keywords: Cell culture | DIGE proteomics | qRT-PCR | Glutathione-S-transferase | Transformation | Transport | Resveratrol | Vitis vinifera
Knowledge Area: Bioquímica y Biología Molecular
Issue Date: 21-Aug-2017
Publisher: Frontiers Media
Citation: Martínez-Márquez A, Martínez-Esteso MJ, Vilella-Antón MT, Sellés-Marchart S, Morante-Carriel JA, Hurtado E, Palazon J and Bru-Martínez R (2017) A Tau Class Glutathione-S-Transferase is Involved in Trans-Resveratrol Transport Out of Grapevine Cells. Front. Plant Sci. 8:1457. doi: 10.3389/fpls.2017.01457
Abstract: Vitis vinifera cell cultures respond to pathogens and elicitors by synthesizing and extracellularly accumulating stilbenoid phytoalexins. Large amounts of trans-resveratrol (t-R) are produced when a cell culture is elicited with methylated cyclodextrins (MBCD), either alone or combined with methyl jasmonate (MeJA). t-R transport to the extracellular medium, which represents the apoplastic space, would place this antifungal defense right in the battlefield to efficiently fight against pathogen attack. Yet despite their physiological relevance, these transport pathways are mostly unknown. A broad hypothesis-free DIGE-based proteomic experiment of a temporal series of elicited grapevine cell cultures was performed to explore the expression profiles of t-R biosynthetic proteins and other co-expressing proteins potentially involved in such a cell response. A correlation between two tau class glutathione-S-transferases (GSTs) with several stilbene synthase and phenylalanine ammonia-lyase isoforms, and with the t-R metabolite itself, was found and further assessed by a qRT-PCR gene expression analysis. The best candidate, GSTU-2, was cloned from the cDNA of the MBCD + MeJA-elicited grapevine cells and used for Agrobacterium-mediated grapevine cell transformation. The non-elicited lines that overexpressed GSTU-2 displayed an extracellular t-R accumulating phenotype, but stabilization of t-R required the addition to culture medium of adsorbent compounds, e.g., PVP or β-cyclodextrin. The wild-type cell cultures accumulated no t-R, not even in the presence of adsorbents. The transient expression of the GSTU-2-GFP fusion proteins in grapevine cells showed localisation in the plasma membrane, and the immunoprecipitation of HA-tagged GSTU-2 revealed its interaction with HIR, a plasma membrane-bound protein. These findings are consistent with a functional role in transport. This is the first report providing several pieces of experimental evidence for the involvement of a specific tau class GST in t-R transport to the extracellular medium.
Sponsor: This work was supported by grants from the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-01, BIO2011-29856-C02-02 and BIO2014-51861-R), The Spanish Ministry of Economy and Competitiveness (PEJ-2014-A-90762/PEJ-2014-P-00289), Generalitat de Catalunya (2014SGR215) and European Funds for Regional Development (FEDER).
URI: http://hdl.handle.net/10045/68852
ISSN: 1664-462X
DOI: 10.3389/fpls.2017.01457
Language: eng
Type: info:eu-repo/semantics/article
Rights: © 2017 Martínez-Márquez, Martínez-Esteso, Vilella-Antón, Sellés-Marchart, Morante-Carriel, Hurtado, Palazon and Bru-Martínez. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Peer Review: si
Publisher version: http://dx.doi.org/10.3389/fpls.2017.01457
Appears in Collections:INV - Proteómica y Genómica Funcional de Plantas - Artículos de Revistas

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