title: Subcellular localization and expression of multiple tomato gamma-aminobutyrate transaminases that utilize both pyruvate and glyoxylate.
abstract: Gamma-aminobutyric acid transaminase (GABA-T) catalyses the breakdown of GABA to succinic semialdehyde. In this report, three GABA-T isoforms were identified in the tomato (Solanum lycopersicum L.) plant. The deduced amino acid sequences of the three isoforms are highly similar over most of their coding regions with the exception of their N-terminal regions. Transient expression of the individual full-length GABA-T isoforms fused to the green fluorescent protein in tobacco suspension-cultured cells revealed their distinct subcellular localizations to the mitochondrion, plastid or cytosol, and that the specific targeting of the mitochondrion- and plastid-localized isoforms is mediated by their predicted N-terminal presequences. Removal of the N-terminal targeting presequences from the mitochondrion and plastid GABA-T isoforms yielded good recovery of the soluble recombinant proteins in Escherichia coli when they were co-expressed with the GroES/EL molecular chaperone complex. Activity assays indicated that all three recombinant isoforms possess both pyruvate- and glyoxylate-dependent GABA-T activities, although the mitochondrial enzyme has a specific activity that is significantly higher than that of its plastid and cytosolic counterparts. Finally, differential expression patterns of the three GABA-T isoforms in reproductive tissues, but not vegetative tissues, suggest unique roles for each enzyme in developmental processes. Overall, these findings, together with recent information about rice and pepper GABA-Ts, indicate that the subcellular distribution of GABA-T in the plant kingdom is highly variable.
authors.affiliation: Department of Plant Agriculture, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
authors.forename: Shawn M
authors.initials: SM
authors.lastname: Clark
authors.forename: Rosa
authors.initials: R
authors.lastname: Di Leo
authors.forename: Owen R
authors.initials: OR
authors.lastname: Van Cauwenberghe
authors.forename: Robert T
authors.initials: RT
authors.lastname: Mullen
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Shelp
mesh_terms: D000612:4-Aminobutyrate Transaminase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D015971:Gene Expression Regulation, Enzymologic; D006038:Glyoxylates / Q000378:metabolism*; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D019289:Pyruvic Acid / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D006038:Glyoxylates; D010940:Plant Proteins; D020033:Protein Isoforms; D019289:Pyruvic Acid; D000612:4-Aminobutyrate Transaminase