title: A coumarin-specific prenyltransferase catalyzes the crucial biosynthetic reaction for furanocoumarin formation in parsley.
abstract: Furanocoumarins constitute a sub-family of coumarin compounds with important defense properties against pathogens and insects, as well as allelopathic functions in plants. Furanocoumarins are divided into two sub-groups according to the alignment of the furan ring with the lactone structure: linear psoralen and angular angelicin derivatives. Determination of furanocoumarin type is based on the prenylation position of the common precursor of all furanocoumarins, umbelliferone, at C6 or C8, which gives rise to the psoralen or angelicin derivatives, respectively. Here, we identified a membrane-bound prenyltransferase PcPT from parsley (Petroselinum crispum), and characterized the properties of the gene product. PcPT expression in various parsley tissues is increased by UV irradiation, with a concomitant increase in furanocoumarin production. This enzyme has strict substrate specificity towards umbelliferone and dimethylallyl diphosphate, and a strong preference for the C6 position of the prenylated product (demethylsuberosin), leading to linear furanocoumarins. The C8-prenylated derivative (osthenol) is also formed, but to a much lesser extent. The PcPT protein is targeted to the plastids in planta. Introduction of this PcPT into the coumarin-producing plant Ruta graveolens showed increased consumption of endogenous umbelliferone. Expression of PcPT and a 4-coumaroyl CoA 2'-hydroxylase gene in Nicotiana benthamiana, which does not produce furanocoumarins, resulted in formation of demethylsuberosin, indicating that furanocoumarin production may be reconstructed by a metabolic engineering approach. The results demonstrate that a single prenyltransferase, such as PcPT, opens the pathway to linear furanocoumarins in parsley, but may also catalyze the synthesis of osthenol, the first intermediate committed to the angular furanocoumarin pathway, in other plants.
authors.affiliation: Laboratoire Agronomie et Environnement Nancy-Colmar, UMR 1121, Université de Lorraine, 2 Avenue de la Forêt de Haye TSA 40602, 54518, Vandoeuvre-lès-Nancy, France; Laboratoire Agronomie et Environnement Nancy-Colmar, UMR 1121, INRA, 2 Avenue de la Forêt de Haye TSA 40602, 54518, Vandoeuvre-lès-Nancy, France.
authors.forename: Fazeelat
authors.initials: F
authors.lastname: Karamat
authors.forename: Alexandre
authors.initials: A
authors.lastname: Olry
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Munakata
authors.forename: Takao
authors.initials: T
authors.lastname: Koeduka
authors.forename: Akifumi
authors.initials: A
authors.lastname: Sugiyama
authors.forename: Cedric
authors.initials: C
authors.lastname: Paris
authors.forename: Alain
authors.initials: A
authors.lastname: Hehn
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bourgaud
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Yazaki
mesh_terms: D001483:Base Sequence; D003374:Coumarins / Q000737:chemistry / Q000378:metabolism; D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism*; D011564:Furocoumarins / Q000737:chemistry / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D019697:Onions / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity; D028528:Petroselinum / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins; D032105:Ruta / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D014466:Ultraviolet Rays; D014468:Umbelliferones / Q000737:chemistry / Q000378:metabolism
chemical_list: D003374:Coumarins; D011564:Furocoumarins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D014468:Umbelliferones; C031477:7-hydroxycoumarin; D004122:Dimethylallyltranstransferase
keywords: Petroselinum crispum; dimethylallyl diphosphate; furanocoumarin; prenyltransferase; umbelliferone