title: CYP703 is an ancient cytochrome P450 in land plants catalyzing in-chain hydroxylation of lauric acid to provide building blocks for sporopollenin synthesis in pollen.
abstract: CYP703 is a cytochrome P450 family specific to land plants. Typically, each plant species contains a single CYP703. Arabidopsis thaliana CYP703A2 is expressed in the anthers of developing flowers. Expression is initiated at the tetrad stage and restricted to microspores and to the tapetum cell layer. Arabidopsis CYP703A2 knockout lines showed impaired pollen development and a partial male-sterile phenotype. Scanning electron and transmission electron microscopy of pollen from the knockout plants showed impaired pollen wall development with absence of exine. The fluorescent layer around the pollen grains ascribed to the presence of phenylpropanoid units in sporopollenin was absent in the CYP703A2 knockout lines. Heterologous expression of CYP703A2 in yeast cells demonstrated that CYP703 catalyzes the conversion of medium-chain saturated fatty acids to the corresponding monohydroxylated fatty acids, with a preferential hydroxylation of lauric acid at the C-7 position. Incubation of recombinant CYP703 with methanol extracts from developing flowers confirmed that lauric acid and in-chain hydroxy lauric acids are the in planta substrate and product, respectively. These data demonstrate that in-chain hydroxy lauric acids are essential building blocks in sporopollenin synthesis and enable the formation of ester and ether linkages with phenylpropanoid units. This study identifies CYP703 as a P450 family specifically involved in pollen development.
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant Biology and Center for Molecular Plant Physiology, Faculty of Life Sciences, University of Copenhagen, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Marc
authors.initials: M
authors.lastname: Morant
authors.forename: Kirsten
authors.initials: K
authors.lastname: Jørgensen
authors.forename: Hubert
authors.initials: H
authors.lastname: Schaller
authors.forename: Franck
authors.initials: F
authors.lastname: Pinot
authors.forename: Birger Lindberg
authors.initials: BL
authors.lastname: Møller
authors.forename: Danièle
authors.initials: D
authors.lastname: Werck-Reichhart
authors.forename: Søren
authors.initials: S
authors.lastname: Bak
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001704:Biopolymers / Q000096:biosynthesis*; D002338:Carotenoids / Q000096:biosynthesis*; D002384:Catalysis; D002473:Cell Wall / Q000648:ultrastructure; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D019143:Evolution, Molecular; D020224:Expressed Sequence Tags; D005298:Fertility; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D006900:Hydroxylation; D007850:Lauric Acids / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D010802:Phylogeny; D011058:Pollen / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D001704:Biopolymers; D007850:Lauric Acids; D012333:RNA, Messenger; C009800:sporopollenin; D002338:Carotenoids; D003577:Cytochrome P-450 Enzyme System; C523532:cytochrome P-450 703A2, Arabidopsis