TitlePurification, molecular cloning, and characterization of glutathione S-transferases (GSTs) from pigmented Vitis vinifera L. cell suspension cultures as putative anthocyanin transport proteins.
Publication TypeJournal Article
Year of Publication2008
AuthorsConn, S, Curtin, CD, Bézier, A, Franco, C, Zhang, W
JournalJ Exp Bot
Volume59
Issue13
Pagination3621-34
Date Published2008
ISSN1460-2431
KeywordsAnthocyanins, Biological Transport, Carrier Proteins, Cells, Cultured, Cloning, Molecular, Gene Expression Regulation, Plant, Genetic Complementation Test, Glutathione Transferase, Models, Molecular, Molecular Sequence Data, Phylogeny, Plant Proteins, Plants, Vacuoles, Vitis, Zea mays
Abstract

The ligandin activity of specific glutathione S-transferases (GSTs) is necessary for the transport of anthocyanins from the cytosol to the plant vacuole. Five GSTs were purified from Vitis vinifera L. cv. Gamay Fréaux cell suspension cultures by glutathione affinity chromatography. These proteins underwent Edman sequencing and mass spectrometry fingerprinting, with the resultant fragments aligned with predicted GSTs within public databases. The corresponding coding sequences were cloned, with heterologous expression in Escherichia coli used to confirm GST activity. Transcriptional profiling of these candidate GST genes and key anthocyanin biosynthetic pathway genes (PAL, CHS, DFR, and UFGT) in cell suspensions and grape berries against anthocyanin accumulation demonstrated strong positive correlation with two sequences, VvGST1 and VvGST4, respectively. The ability of VvGST1 and VvGST4 to transport anthocyanins was confirmed in the heterologous maize bronze-2 complementation model, providing further evidence for their function as anthocyanin transport proteins in grape cells. Furthermore, the differential induction of VvGST1 and VvGST4 in suspension cells and grape berries suggests functional differences between these two proteins. Further investigation of these candidate ligandins may identify a mechanism for manipulating anthocyanin accumulation in planta and in vitro suspension cells.

DOI10.1093/jxb/ern217
Alternate JournalJ. Exp. Bot.
PubMed ID18836188
PubMed Central IDPMC2561157