title: Functional expression of PHO1 to the Golgi and trans-Golgi network and its role in export of inorganic phosphate.
abstract: Arabidopsis thaliana PHO1 is primarily expressed in the root vascular cylinder and is involved in the transfer of inorganic phosphate (Pi) from roots to shoots. To analyze the role of PHO1 in transport of Pi, we have generated transgenic plants expressing PHO1 in ectopic A. thaliana tissues using an estradiol-inducible promoter. Leaves treated with estradiol showed strong PHO1 expression, leading to detectable accumulation of PHO1 protein. Estradiol-mediated induction of PHO1 in leaves from soil-grown plants, in leaves and roots of plants grown in liquid culture, or in leaf mesophyll protoplasts, was all accompanied by the specific release of Pi to the extracellular medium as early as 2-3 h after addition of estradiol. Net Pi export triggered by PHO1 induction was enhanced by high extracellular Pi and weakly inhibited by the proton-ionophore carbonyl cyanide m-chlorophenylhydrazone. Expression of a PHO1-GFP construct complementing the pho1 mutant revealed GFP expression in punctate structures in the pericycle cells but no fluorescence at the plasma membrane. When expressed in onion epidermal cells or in tobacco mesophyll cells, PHO1-GFP was associated with similar punctate structures that co-localized with the Golgi/trans-Golgi network and uncharacterized vesicles. However, PHO1-GFP could be partially relocated to the plasma membrane in leaves infiltrated with a high-phosphate solution. Together, these results show that PHO1 can trigger Pi export in ectopic plant cells, strongly indicating that PHO1 is itself a Pi exporter. Interestingly, PHO1-mediated Pi export was associated with its localization to the Golgi and trans-Golgi networks, revealing a role for these organelles in Pi transport.
authors.affiliation: Department of Plant Molecular Biology, University of Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: A Bulak
authors.initials: AB
authors.lastname: Arpat
authors.forename: Pasqualina
authors.initials: P
authors.lastname: Magliano
authors.forename: Stefanie
authors.initials: S
authors.lastname: Wege
authors.forename: Hatem
authors.initials: H
authors.lastname: Rouached
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Stefanovic
authors.forename: Yves
authors.initials: Y
authors.lastname: Poirier
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D006056:Golgi Apparatus / Q000378:metabolism*; D006706:Homeostasis; D009566:Nitrates / Q000378:metabolism; D019697:Onions / Q000235:genetics / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D018520:Plant Shoots / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011523:Protoplasts; D011993:Recombinant Fusion Proteins; D036226:Seedlings / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D009566:Nitrates; C460111:PHO1 protein, Arabidopsis; D010710:Phosphates; D011993:Recombinant Fusion Proteins