title: Dimerization of N-methyltransferases involved in caffeine biosynthesis.
abstract: Caffeine is synthesized from the precursor xanthosine through three methylation and one nucleoside removal steps. Methylation is catalyzed by N-methyltransferases, designated as CaXMT1, CaMXMT1 and CaDXMT1, which, respectively, convert xanthosine into 7-methylxanthosine, 7-methylxanthine into 3,7-dimethylxanthine, and 3,7-dimethylxanthine into 1,3,7-trimethylxanthine (caffeine). In the present study, we examined their cytological and biochemical properties using fusion proteins with fluorescent proteins. All three enzymes were found to localize in cytosol as visualized by green fluorescence protein fusions. The possibility of dimer formation among these enzyme proteins was examined in vivo by transient expression of bimolecular fluorescence complementation of yellow fluorescent protein (YFP) using onion epidermal cell layers. Results showed that each enzyme protein formed a homo-dimer in cytosol as seen by a clear reconstituted YFP fluorescence. In addition, each enzyme also formed a hetero-dimer with each of the other two enzymes in cytosol. The biological significance of dimerization among structurally resembling methyltransferases involved in caffeine biosynthesis is discussed.
authors.affiliation: Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Nara, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kodama
authors.forename: T
authors.initials: T
authors.lastname: Shinya
authors.forename: H
authors.initials: H
authors.lastname: Sano
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D002110:Caffeine / Q000096:biosynthesis*; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D008164:Luminescent Proteins / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000032:analysis / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D018076:DNA, Complementary; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D002110:Caffeine; D008780:Methyltransferases