title: Proton-dependent glutamine uptake by aphid bacteriocyte amino acid transporter ApGLNT1.
abstract: Aphids house large populations of the gammaproteobacterial symbiont Buchnera aphidicola in specialized bacteriocyte cells. The combined biosynthetic capability of the holobiont (Acyrthosiphon pisum and Buchnera) is sufficient for biosynthesis of all twenty protein coding amino acids, including amino acids that animals alone cannot synthesize; and that are present at low concentrations in A. pisum's plant phloem sap diet. Collaborative holobiont amino acid biosynthesis depends on glutamine import into bacteriocytes, which serves as a nitrogen-rich amino donor for biosynthesis of other amino acids. Recently, we characterized A. pisum glutamine transporter 1 (ApGLNT1), a member of the amino acid/auxin permease family, as the dominant bacteriocyte plasma membrane glutamine transporter. Here we show ApGLNT1 to be structurally and functionally related to mammalian proton-dependent amino acid transporters (PATs 1-4). Using functional expression in Xenopus laevis oocytes, combined with two-electrode voltage clamp electrophysiology we demonstrate that ApGLNT1 is electrogenic and that glutamine induces large inward currents. ApGLNT1 glutamine induced currents are dependent on external glutamine concentration, proton (H+) gradient across the membrane, and membrane potential. Based on these transport properties, ApGLNT1-mediated glutamine uptake into A. pisum bacteriocytes can be regulated by changes in either proton gradients across the plasma membrane or membrane potential.
authors.affiliation: Department of Biology, University of Miami, Coral Gables, FL 33146, USA. Electronic address: dan.price@bio.miami.edu.
authors.forename: Daniel R G
authors.initials: DR
authors.lastname: Price
authors.affiliation: Department of Biology, University of Miami, Coral Gables, FL 33146, USA. Electronic address: acwilson@bio.miami.edu.
authors.forename: Alex C C
authors.initials: AC
authors.lastname: Wilson
authors.affiliation: Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami, Miami, FL 33136, USA. Electronic address: cluetje@med.miami.edu.
authors.forename: Charles W
authors.initials: CW
authors.lastname: Luetje
mesh_terms: D000818:Animals; D001042:Aphids / Q000382:microbiology*; D020635:Buchnera / Q000378:metabolism*; D002478:Cells, Cultured; D005973:Glutamine / Q000493:pharmacokinetics*; D015640:Ion Channel Gating / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D011522:Protons; D014982:Xenopus laevis
chemical_list: D011522:Protons; D005973:Glutamine
keywords: Amino acid/auxin permease; Electrogenic transport; Holobiont; Symbiosis