title: Gene network reconstruction identifies the authentic trans-prenyl diphosphate synthase that makes the solanesyl moiety of ubiquinone-9 in Arabidopsis.
abstract: Ubiquinone (coenzyme Q) is the generic name of a class of lipid-soluble electron carriers formed of a redox active benzoquinone ring attached to a prenyl side chain. The length of the latter varies among species, and depends upon the product specificity of a trans-long-chain prenyl diphosphate synthase that elongates an allylic diphosphate precursor. In Arabidopsis, this enzyme is assumed to correspond to an endoplasmic reticulum-located solanesyl diphosphate synthase, although direct genetic evidence was lacking. In this study, the reconstruction of the functional network of Arabidopsis genes linked to ubiquinone biosynthesis singled out an unsuspected solanesyl diphosphate synthase candidate--product of gene At2g34630--that, extraordinarily, had been shown previously to be targeted to plastids and to contribute to the biosynthesis of gibberellins. Green fluorescent protein (GFP) fusion experiments in tobacco and Arabidopsis, and complementation of a yeast coq1 knockout lacking mitochondrial hexaprenyl diphosphate synthase demonstrated that At2g34630 is also targeted to mitochondria. At2g34630 is the main--if not sole--contributor to solanesyl diphosphate synthase activity required for the biosynthesis of ubiquinone, as demonstrated by the dramatic (75-80%) reduction of the ubiquinone pool size in corresponding RNAi lines. Overexpression of At2g34630 gave up to a 40% increase in ubiquinone content compared to wild-type plants. None of the silenced or overexpressing lines, in contrast, displayed altered levels of plastoquinone. Phylogenetic analyses revealed that At2g34630 is the only Arabidopsis trans-long-chain prenyl diphosphate synthase that clusters with the Coq1 orthologs involved in the biosynthesis of ubiquinone in other eukaryotes.
authors.affiliation: Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
authors.forename: Anne-Lise
authors.initials: AL
authors.lastname: Ducluzeau
authors.forename: Yashitola
authors.initials: Y
authors.lastname: Wamboldt
authors.forename: Christian G
authors.initials: CG
authors.lastname: Elowsky
authors.forename: Sally A
authors.initials: SA
authors.lastname: Mackenzie
authors.forename: Robert C
authors.initials: RC
authors.lastname: Schuurink
authors.forename: Gilles J C
authors.initials: GJ
authors.lastname: Basset
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D002736:Chloroplasts / Q000201:enzymology; D003001:Cloning, Molecular; D055786:Gene Knockout Techniques; D053263:Gene Regulatory Networks / Q000235:genetics*; D005816:Genetic Complementation Test; D049452:Green Fluorescent Proteins; D008928:Mitochondria / Q000201:enzymology; D009154:Mutation; D010802:Phylogeny; D030821:Plants, Genetically Modified; D010971:Plastoquinone / Q000378:metabolism; D034622:RNA Interference; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism; D014451:Ubiquinone / Q000737:chemistry / Q000378:metabolism*
chemical_list: D013729:Terpenes; D014451:Ubiquinone; D049452:Green Fluorescent Proteins; D019883:Alkyl and Aryl Transferases; C022644:trans-octaprenyltranstransferase; C030571:ubiquinone 9; D010971:Plastoquinone