ABCC1, an ATP Binding Cassette Protein from Grape Berry, Transports Anthocyanidin 3-O-Glucosides

Abstract

ORCID ID: 0000-0002-9663-5371 (RN).Accumulation of anthocyanins in the exocarp of red grapevine (Vitis vinifera) cultivars is one of several events that characterize the onset of grape berry ripening (véraison). Despite our thorough understanding of anthocyanin biosynthesis and regulation, little is known about the molecular aspects of their transport. The participation of ATP binding cassette (ABC) proteins in vacuolar anthocyanin transport has long been a matter of debate. Here, we present biochemical evidence that an ABC protein, ABCC1, localizes to the tonoplast and is involved in the transport of glucosylated anthocyanidins. ABCC1 is expressed in the exocarp throughout berry development and ripening, with a significant increase at véraison (i.e., the onset of ripening). Transport experiments using microsomes isolated from ABCC1-expressing yeast cells showed that ABCC1 transports malvidin 3-Oglucoside. The transport strictly depends on the presence of GSH, which is cotransported with the anthocyanins and is sensitive to inhibitors of ABC proteins. By exposing anthocyanin-producing grapevine root cultures to buthionine sulphoximine, which reduced GSH levels, a decrease in anthocyanin concentration is observed. In conclusion, we provide evidence that ABCC1 acts as an anthocyanin transporter that depends on GSH without the formation of an anthocyanin-GSH conjugate.

Anthocyanin Vacuolar Inclusions Form by a Microautophagy Mechanism

“…According to the ligandin model, cytoplasmic anthocyanins bind to specific glutathione S-transferases (GSTs), encoded in Arabidopsis by TRANSPARENT TESTA19 (TT19), in maize (Zea mays) by BRONZE2, and in petunia (Petunia hybrida) by AN9 (Marrs et al, 1995;Alfenito et al, 1998;Kitamura et al, 2004;Conn et al, 2008;Sun et al, 2012). These GSTs escort anthocyanins to the vacuolar membrane or tonoplast where some transporters of the ABC (ATP-binding cassette) and MATE (multidrug and toxin extrusion) families transfer anthocyanin molecules into the vacuolar lumen (Goodman et al, 2004;Marinova et al, 2007;Gomez et al, 2009;Francisco et al, 2013). The vesicular transport model postulates that anthocyanins enter the ER lumen and are transported in vesicles and/or membrane-bound organelles to the vacuole.…”

Vacuolar Transport of Abscisic Acid Glucosyl Ester Is Mediated by ATP-Binding Cassette and Proton-Antiport Mechanisms in Arabidopsis

“…The energized transport of glucosides of secondary metabolites and xenobiotics into plant vacuoles is well documented. The anthocyanin malvidin-3-O-glucoside is transported into vacuoles of grape (Vitis vinifera) berries by the ABCC transporter ABCC1 from grape (Francisco et al, 2013). Proton gradient-dependent vacuolar transport mechanisms were reported for diverse flavonoid glucosides (Klein et al, 1996;Frangne et al, 2002;Zhao and Dixon, 2009;Zhao et al, 2011).…”

“…The vacuolar membrane localization of Arabidopsis ABCC-type transporters and the recent demonstration that grape ABCC1 mediates the vacuolar transport of anthocyanidin glucosides (Kang et al, 2011;Francisco et al, 2013) suggested the participation of ABCC-type transporters in vacuolar ABA-GE accumulation. The Arabidopsis AtABCC1 and especially AtABCC2 mediate the transport of structurally diverse metabolites, such as phytochelatins, folates, and conjugates of chlorophyll catabolite and xenobiotics (Liu et al, 2001;Frelet-Barrand et al, 2008;Raichaudhuri et al, 2009;Song et al, 2010).…”

Flavonoid transport mechanisms: how to go, and with whom

“…Three distinct but potentially nonexclusive mechanisms for flavonoid transport in plant cells have been proposed: vesicle trafficking, membrane transporter, and GST mediated [2]. Over the past 5 years, significant progress has been made in understanding the mechanisms for the vacuolar sequestration of flavonoids, including the functional characterization of new flavonoid transporters with novel biochemical properties and in planta biological functions [3][4][5]. Long-standing questions regarding the mechanisms by which ATP-binding cassette (ABC) transporters and GSTs function in flavonoid transport have been partly addressed [5][6][7][8].…”