title: Characterization and subcellular localization of geranylgeranyl diphosphate synthase from Catharanthus roseus.
abstract: The enzyme geranylgeranyl diphosphate synthase (GGPS: EC 2.5.1.1, EC 2.5.1.10, EC 2.5.1.29) catalyses the formation of geranylgeranyl diphosphate (GGPP) from isopentenyl diphosphate and dimethylallyl diphosphate via three successive condensation reactions. A full-length nucleotide sequence of GGPS (named CrGGPS) was cloned from the medicinal plant Catharanthus roseus. The deduced polypeptide has 383 amino acids with a calculated mass of 41.6 kDa and possesses prenyltransferase signatures characteristic of plant type II GGPS. The enzyme was characterized by functional complementation in carotenoid accumulating strains of Escherichia coli. When cultures of Catharanthus cell lines were treated with methyljasmonate, no specific increase in transcript levels were observed. In plants, GGPS are encoded by a small multigene family and the isoforms have been shown to be localized in three different subcellular compartments: chloroplast, endoplasmic reticulum and mitochondria. We investigated the subcellular distribution of CrGGPS through transient transformations of C. roseus cells with a yellow fluorescent protein-fused construct. Our results clearly indicate that CrGGPS is located to plastids within stroma and stromules.
authors.affiliation: EA2106, Biomolécules et Biotechnologies Végétales, Université François-Rabelais, 31 avenue Monge, 37200 Tours, France.
authors.forename: Insaf
authors.initials: I
authors.lastname: Thabet
authors.forename: Grégory
authors.initials: G
authors.lastname: Guirimand
authors.forename: Anthony
authors.initials: A
authors.lastname: Guihur
authors.forename: Arnaud
authors.initials: A
authors.lastname: Lanoue
authors.forename: Vincent
authors.initials: V
authors.lastname: Courdavault
authors.forename: Nicolas
authors.initials: N
authors.lastname: Papon
authors.forename: Sadok
authors.initials: S
authors.lastname: Bouzid
authors.forename: Nathalie
authors.initials: N
authors.lastname: Giglioli-Guivarc'h
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Simkin
authors.forename: Marc
authors.initials: M
authors.lastname: Clastre
mesh_terms: D000085:Acetates; D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D030008:Catharanthus / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003517:Cyclopentanes; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis; D004926:Escherichia coli; D051231:Farnesyltranstransferase / Q000235:genetics* / Q000378:metabolism; D005816:Genetic Complementation Test; D042541:Intracellular Space / Q000378:metabolism; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D054883:Oxylipins; D018087:Plastids / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D000085:Acetates; D001426:Bacterial Proteins; D003517:Cyclopentanes; D017931:DNA Primers; D018076:DNA, Complementary; D008164:Luminescent Proteins; D054883:Oxylipins; C054562:yellow fluorescent protein, Bacteria; C072239:methyl jasmonate; D051231:Farnesyltranstransferase