title: PIP2 regulates psychostimulant behaviors through its interaction with a membrane protein.
abstract: Phosphatidylinositol (4,5)-bisphosphate (PIP2) regulates the function of ion channels and transporters. Here, we demonstrate that PIP2 directly binds the human dopamine (DA) transporter (hDAT), a key regulator of DA homeostasis and a target of the psychostimulant amphetamine (AMPH). This binding occurs through electrostatic interactions with positively charged hDAT N-terminal residues and is shown to facilitate AMPH-induced, DAT-mediated DA efflux and the psychomotor properties of AMPH. Substitution of these residues with uncharged amino acids reduces hDAT-PIP2 interactions and AMPH-induced DA efflux without altering the hDAT physiological function of DA uptake. We evaluated the significance of this interaction in vivo using locomotion as a behavioral assay in Drosophila melanogaster. Expression of mutated hDAT with reduced PIP2 interaction in Drosophila DA neurons impairs AMPH-induced locomotion without altering basal locomotion. We present what is to our knowledge the first demonstration of how PIP2 interactions with a membrane protein can regulate the behaviors of complex organisms.
authors.affiliation: Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.|Neuroscience Program in Substance Abuse, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Hamilton
authors.affiliation: Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.
authors.forename: Andrea N
authors.initials: AN
authors.lastname: Belovich
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College of Cornell University, New York, New York 10065, United States.
authors.forename: George
authors.initials: G
authors.lastname: Khelashvili
authors.affiliation: Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.
authors.forename: Christine
authors.initials: C
authors.lastname: Saunders
authors.affiliation: Department of Molecular Physiology & Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.|Neuroscience Program in Substance Abuse, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.
authors.forename: Kevin
authors.initials: K
authors.lastname: Erreger
authors.affiliation: Department of Psychiatry, Columbia University College of Physicians & Surgeons, New York, New York 10032, United States.|Department of Pharmacology, Columbia University College of Physicians & Surgeons, New York, New York 10032, United States.
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Javitch
authors.affiliation: Institute of Pharmacology, Center for Physiology and Pharmacology, Medical University Vienna, Vienna, Austria.
authors.forename: Harald H
authors.initials: HH
authors.lastname: Sitte
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College of Cornell University, New York, New York 10065, United States.|HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute of Computational Biomedicine, Weill Cornell Medical College of Cornell University, New York, New York 10065, United States.
authors.forename: Harel
authors.initials: H
authors.lastname: Weinstein
authors.affiliation: Department of Molecular Physiology & Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.|Neuroscience Program in Substance Abuse, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.
authors.forename: Heinrich J G
authors.initials: HJG
authors.lastname: Matthies
authors.affiliation: Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.|Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.|Department of Molecular Physiology & Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.|Neuroscience Program in Substance Abuse, Vanderbilt University School of Medicine, Nashville, TN 37232-8548, United States.
authors.forename: Aurelio
authors.initials: A
authors.lastname: Galli
mesh_terms: D019943:Amino Acid Substitution; D000661:Amphetamine / Q000494:pharmacology*; D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects; D000697:Central Nervous System Stimulants / Q000494:pharmacology*; D004298:Dopamine / Q000378:metabolism; D050483:Dopamine Plasma Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000187:drug effects* / Q000502:physiology; D015870:Gene Expression; D006801:Humans; D008124:Locomotion / Q000187:drug effects; D008958:Models, Molecular; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism* / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D019076:Transgenes
chemical_list: D000697:Central Nervous System Stimulants; D050483:Dopamine Plasma Membrane Transport Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D000661:Amphetamine; D004298:Dopamine