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PLoS One

Title:		Involvement of the leaf-specific multidrug and toxic compound extrusion (MATE) transporter Nt-JAT2 in vacuolar sequestration of nicotine in Nicotiana tabacum
Author(s):		Shitan N; Minami S; Morita M; Hayashida M; Ito S; Takanashi K; Omote H; Moriyama Y; Sugiyama A; Goossens A; Moriyasu M; Yazaki K;
Address:		"Department of Natural Medicinal Chemistry, Kobe Pharmaceutical University, Kobe, Hyogo, Japan; Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Kyoto, Japan. Department of Natural Medicinal Chemistry, Kobe Pharmaceutical University, Kobe, Hyogo, Japan. Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Kyoto, Japan. Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan. Department of Plant Systems Biology, VIB, Gent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, Gent, Belgium"
Journal Title:		PLoS One
Year:		2014
Volume:		20140930
Issue:		9
Page Number:		e108789 -
DOI:		10.1371/journal.pone.0108789
ISSN/ISBN:		1932-6203 (Electronic) 1932-6203 (Linking)
Abstract:		"Alkaloids play a key role in higher plant defense against pathogens and herbivores. Following its biosynthesis in root tissues, nicotine, the major alkaloid of Nicotiana species, is translocated via xylem transport toward the accumulation sites, leaf vacuoles. Our transcriptome analysis of methyl jasmonate-treated tobacco BY-2 cells identified several multidrug and toxic compound extrusion (MATE) transporter genes. In this study, we characterized a MATE gene, Nicotiana tabacum jasmonate-inducible alkaloid transporter 2 (Nt-JAT2), which encodes a protein that has 32% amino acid identity with Nt-JAT1. Nt-JAT2 mRNA is expressed at a very low steady state level in whole plants, but is rapidly upregulated by methyl jasmonate treatment in a leaf-specific manner. To characterize the function of Nt-JAT2, yeast cells were used as the host organism in a cellular transport assay. Nt-JAT2 was localized at the plasma membrane in yeast cells. When incubated in nicotine-containing medium, the nicotine content in Nt-JAT2-expressing cells was significantly lower than in control yeast. Nt-JAT2-expressing cells also showed lower content of other alkaloids like anabasine and anatabine, but not of flavonoids, suggesting that Nt-JAT2 transports various alkaloids including nicotine. Fluorescence assays in BY-2 cells showed that Nt-JAT2-GFP was localized to the tonoplast. These findings indicate that Nt-JAT2 is involved in nicotine sequestration in leaf vacuoles following the translocation of nicotine from root tissues"
Keywords:		Acetates/pharmacology Alkaloids/metabolism Anabasine/metabolism Cell Membrane/metabolism Cyclopentanes/pharmacology Flavonoids/metabolism Gene Expression Profiling Nicotine/*metabolism/pharmacology Oxylipins/pharmacology Phylogeny Plant Cells/drug effects;
Notes:		"MedlineShitan, Nobukazu Minami, Shota Morita, Masahiko Hayashida, Minaho Ito, Shingo Takanashi, Kojiro Omote, Hiroshi Moriyama, Yoshinori Sugiyama, Akifumi Goossens, Alain Moriyasu, Masataka Yazaki, Kazufumi eng Research Support, Non-U.S. Gov't 2014/10/01 PLoS One. 2014 Sep 30; 9(9):e108789. doi: 10.1371/journal.pone.0108789. eCollection 2014"