title: Targeting of a Nicotiana plumbaginifolia H+ -ATPase to the plasma membrane is not by default and requires cytosolic structural determinants.
abstract: The structural determinants involved in the targeting of multitransmembrane-span proteins to the plasma membrane (PM) remain poorly understood. The plasma membrane H+ -ATPase (PMA) from Nicotiana plumbaginifolia, a well-characterized 10 transmembrane-span enzyme, was used as a model to identify structural elements essential for targeting to the PM. When PMA2 and PMA4, representatives of the two main PMA subfamilies, were fused to green fluorescent protein (GFP), the chimeras were shown to be still functional and to be correctly and rapidly targeted to the PM in transgenic tobacco. By contrast, chimeric proteins containing various combinations of PMA transmembrane spanning domains accumulated in the Golgi apparatus and not in the PM and displayed slow traffic properties through the secretory pathway. Individual deletion of three of the four cytosolic domains did not prevent PM targeting, but deletion of the large loop or of its nucleotide binding domain resulted in GFP fluorescence accumulating exclusively in the endoplasmic reticulum. The results show that, at least for this polytopic protein, the PM is not the default pathway and that, in contrast with single-pass membrane proteins, cytosolic structural determinants are required for correct targeting.
authors.affiliation: Unité de Biochimie Physiologique, Institut des Sciences de la Vie, Université Catholique de Louvain, B-1348 Louvain-la-Neuve, Belgium.
authors.forename: Benoit
authors.initials: B
authors.lastname: Lefebvre
authors.forename: Henri
authors.initials: H
authors.lastname: Batoko
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Duby
authors.forename: Marc
authors.initials: M
authors.lastname: Boutry
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D021381:Protein Transport; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D011523:Protoplasts / Q000166:cytology; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014026:Nicotiana / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D006180:Proton-Translocating ATPases