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SYSTEMS BIOLOGY, MOLECULAR BIOLOGY, AND GENE REGULATION

Grapevine MATE-Type Proteins Act as Vacuolar H⁺-Dependent Acylated Anthocyanin Transporters^1,[W],[OA]

UMR Sciences pour l'nologie, INRA Campus SupAgro, F–34060 Montpellier, France (C.G., N.T., S.V., C.V., J.-M.S., J.-P.M., V.C., A.A.); UMR Diversité et Adaptation des Plantes Cultivées 1097, INRA Campus SupAgro, F–34060 Montpellier, France (L.T., A.F.-L.); and Zurich Basel Plant Science Center, University of Zurich, Plant Biology, CH–8008 Zurich, Switzerland (M.K.)

In grapevine (Vitis vinifera), anthocyanins are responsible for most of the red, blue, and purple pigmentation found in the skin of berries. In cells, anthocyanins are synthesized in the cytoplasm and accumulated into the vacuole. However, little is known about the transport of these compounds through the tonoplast. Recently, the sequencing of the grapevine genome allowed us to identify genes encoding proteins with high sequence similarity to the Multidrug And Toxic Extrusion (MATE) family. Among them, we selected two genes as anthocyanin transporter candidates and named them anthoMATE1 (AM1) and AM3. The expression of both genes was mainly fruit specific and concomitant with the accumulation of anthocyanin pigment. Subcellular localization assays in grapevine hairy roots stably transformed with AM1:: or AM3::green fluorescent protein fusion protein revealed that AM1 and AM3 are primarily localized to the tonoplast. Yeast vesicles expressing anthoMATEs transported acylated anthocyanins in the presence of MgATP. Inhibitor studies demonstrated that AM1 and AM3 proteins act in vitro as vacuolar H⁺-dependent acylated anthocyanin transporters. By contrast, under our experimental conditions, anthoMATEs could not transport malvidin 3-O-glucoside or cyanidin 3-O-glucoside, suggesting that the acyl conjugation was essential for the uptake. Taken together, these results provide evidence that in vitro the two grapevine AM1 and AM3 proteins mediate specifically acylated anthocyanin transport.

² Present address: Philip Morris International, R&D, Quai Jeanrenaud 56, 2000 Neuchâtel, Switzerland.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Agnès Ageorges (ageorges{at}supagro.inra.fr).

^[W] The online version of this article contains Web-only data.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.109.135624

^* Corresponding author; e-mail ageorges{at}supagro.inra.fr.

Received January 13, 2009; accepted March 16, 2009; published March 18, 2009.

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