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Mutation of Tyrosine 470 of Human Dopamine Transporter is Critical for HIV-1 Tat-Induced Inhibition of Dopamine Transport and Transporter Conformational Transitions

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Abstract

HIV-1 Tat protein plays a crucial role in perturbations of the dopamine (DA) system. Our previous studies have demonstrated that Tat decreases DA uptake, and allosterically modulates DA transporter (DAT) function. In the present study, we have found that Tat interacts directly with DAT, leading to inhibition of DAT function. Through computational modeling and simulations, a potential recognition binding site of human DAT (hDAT) for Tat was predicted. Mutation of tyrosine470 (Y470H) attenuated Tat-induced inhibition of DA transport, implicating the functional relevance of this residue for Tat binding to hDAT. Y470H reduced the maximal velocity of [3H]DA uptake without changes in the Km and IC50 values for DA inhibition of DA uptake but increased DA uptake potency for cocaine and GBR12909, suggesting that this residue does not overlap with the binding sites in hDAT for substrate but is critical for these inhibitors. Furthermore, Y470H also led to transporter conformational transitions by affecting zinc modulation of DA uptake and WIN35,428 binding as well as enhancing basal DA efflux. Collectively, these findings demonstrate Tyr470 as a functional recognition residue in hDAT for Tat-induced inhibition of DA transport and transporter conformational transitions. The consequence of mutation at this residue is to block the functional binding of Tat to hDAT without affecting physiological DA transport.

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References

  • Aksenov MY, Aksenova MV, Mactutus CF, Booze RM (2009) Attenuated neurotoxicity of the transactivation-defective HIV-1 Tat protein in hippocampal cell cultures. Exp Neurol 219:586–590

    Article  PubMed  CAS  Google Scholar 

  • Albini A, Ferrini S, Benelli R, Sforzini S, Giunciuglio D, Aluigi MG, Proudfoot AE, Alouani S, Wells TN, Mariani G, Rabin RL, Farber JM, Noonan DM (1998) HIV-1 Tat protein mimicry of chemokines. Proc Natl Acad Sci U S A 95:13153–13158

    Article  PubMed  CAS  Google Scholar 

  • Andersen PH, Jansen JA, Nielsen EB (1987) [3H]GBR 12935 binding in vivo in mouse brain: Labelling of a piperazine acceptor site. Eur J Pharmacol 144:1–6

    Article  PubMed  CAS  Google Scholar 

  • Berger JR, Arendt G (2000) HIV dementia: The role of the basal ganglia and dopaminergic systems. J Psychopharmacol 14:214–221

    Article  PubMed  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Buch S, Yao H, Guo M, Mori T, Su TP, Wang J (2011) Cocaine and HIV-1 interplay: Molecular mechanisms of action and addiction. J Neuroimmune Pharmacol 6:503–515

    Article  PubMed  Google Scholar 

  • Chang L, Wang GJ, Volkow ND, Ernst T, Telang F, Logan J, Fowler JS (2008) Decreased brain dopamine transporters are related to cognitive deficits in HIV patients with or without cocaine abuse. NeuroImage 42:869–878

    Article  PubMed  Google Scholar 

  • Chen N, Rickey J, Berfield JL, Reith ME (2004) Aspartate 345 of the dopamine transporter is critical for conformational changes in substrate translocation and cocaine binding. J Biol Chem 279:5508–5519

    Article  PubMed  CAS  Google Scholar 

  • Del Valle L, Croul S, Morgello S, Amini S, Rappaport J, Khalili K (2000) Detection of HIV-1 Tat and JCV capsid protein, VP1, in AIDS brain with progressive multifocal leukoencephalopathy. J Neurovirol 6:221–228

    Article  PubMed  Google Scholar 

  • Ernst T, Yakupov R, Nakama H, Crocket G, Cole M, Watters M, Ricardo-Dukelow ML, Chang L (2009) Declined neural efficiency in cognitively stable human immunodeficiency virus patients. Ann Neurol 65:316–325

    Article  PubMed  Google Scholar 

  • Ferris MJ, Mactutus CF, Booze RM (2008) Neurotoxic profiles of HIV, psychostimulant drugs of abuse, and their concerted effect on the brain: Current status of dopamine system vulnerability in NeuroAIDS. Neurosci Biobehav Rev 32:883–909

    Article  PubMed  CAS  Google Scholar 

  • Ferris MJ, Frederick-Duus D, Fadel J, Mactutus CF, Booze RM (2010) Hyperdopaminergic tone in HIV-1 protein treated rats and cocaine sensitization. J Neurochem 115:885–896

    Article  PubMed  CAS  Google Scholar 

  • Gabdoulline RR, Wade RC (1998) Brownian dynamics simulation of protein-protein diffusional encounter. Methods 14:329–341

    Article  PubMed  CAS  Google Scholar 

  • Gannon P, Khan MZ, Kolson DL (2011) Current understanding of HIV-associated neurocognitive disorders pathogenesis. Curr Opin Neurol 24:275–283

    Article  PubMed  Google Scholar 

  • Gaskill PJ, Calderon TM, Luers AJ, Eugenin EA, Javitch JA, Berman JW (2009) Human immunodeficiency virus (HIV) infection of human macrophages is increased by dopamine: A bridge between HIV-associated neurologic disorders and drug abuse. Am J Pathol 175:1148–1159

    Article  PubMed  CAS  Google Scholar 

  • Gelman BB, Lisinicchia JG, Chen T, Johnson KM, Jennings K, Freeman DH Jr, Soukup VM (2012) Prefrontal dopaminergic and enkephalinergic synaptic accommodation in HIV-associated neurocognitive disorders and encephalitis. J Neuroimmune Pharmacol 7:686–700

    Article  PubMed  Google Scholar 

  • Guptaroy B, Fraser R, Desai A, Zhang M, Gnegy ME (2011) Site-directed mutations near transmembrane domain 1 alter conformation and function of norepinephrine and dopamine transporters. Mol Pharmacol 79:520–532

    Article  PubMed  CAS  Google Scholar 

  • Guptaroy B, Zhang M, Bowton E, Binda F, Shi L, Weinstein H, Galli A, Javitch JA, Neubig RR, Gnegy ME (2009) A juxtamembrane mutation in the N terminus of the dopamine transporter induces preference for an inward-facing conformation. Mol Pharmacol 75:514–524

    Article  PubMed  CAS  Google Scholar 

  • Harrod SB, Mactutus CF, Fitting S, Hasselrot U, Booze RM (2008) Intra-accumbal Tat1-72 alters acute and sensitized responses to cocaine. Pharmacol Biochem Behav 90:723–729

    Article  PubMed  CAS  Google Scholar 

  • Ho BK, Gruswitz F (2008) HOLLOW: generating accurate representations of channel and interior surfaces in molecular structures. BMC structural biology 8:49.

  • Huang X, Zhan CG (2007) How dopamine transporter interacts with dopamine: Insights from molecular modeling and simulation. Biophys J 93:3627–3639

    Article  PubMed  CAS  Google Scholar 

  • Huang X, Gu HH, Zhan CG (2009) Mechanism for cocaine blocking the transport of dopamine: Insights from molecular modeling and dynamics simulations. J Phys Chem B 113:15057–15066

    Article  PubMed  CAS  Google Scholar 

  • Hudson L, Liu J, Nath A, Jones M, Raghavan R, Narayan O, Male D, Everall I (2000) Detection of the human immunodeficiency virus regulatory protein tat in CNS tissues. J Neurovirol 6:145–155

    Article  PubMed  CAS  Google Scholar 

  • Kumar AM, Ownby RL, Waldrop-Valverde D, Fernandez B, Kumar M (2011) Human immunodeficiency virus infection in the CNS and decreased dopamine availability: Relationship with neuropsychological performance. J Neurovirol 17:26–40

    Article  PubMed  CAS  Google Scholar 

  • Kumar AM, Fernandez JB, Singer EJ, Commins D, Waldrop-Valverde D, Ownby RL, Kumar M (2009) Human immunodeficiency virus type 1 in the central nervous system leads to decreased dopamine in different regions of postmortem human brains. J Neurovirol 15:257–274

    Article  PubMed  CAS  Google Scholar 

  • Lamers SL, Salemi M, Galligan DC, Morris A, Gray R, Fogel G, Zhao L, McGrath MS (2010) Human immunodeficiency virus-1 evolutionary patterns associated with pathogenic processes in the brain. J Neurovirol 16:230–241

    Article  PubMed  CAS  Google Scholar 

  • Li W, Li G, Steiner J, Nath A (2009) Role of Tat protein in HIV neuropathogenesis. Neurotox Res 16:205–220

    Article  PubMed  CAS  Google Scholar 

  • Li W, Huang Y, Reid R, Steiner J, Malpica-Llanos T, Darden TA, Shankar SK, Mahadevan A, Satishchandra P, Nath A (2008) NMDA receptor activation by HIV-Tat protein is clade dependent. J Neurosci 28:12190–12198

    Article  PubMed  CAS  Google Scholar 

  • Loland CJ, Norgaard-Nielsen K, Gether U (2003) Probing dopamine transporter structure and function by Zn2 + −site engineering. Eur J Pharmacol 479:187–197

    Article  PubMed  CAS  Google Scholar 

  • Loland CJ, Norregaard L, Litman T, Gether U (2002) Generation of an activating Zn(2+) switch in the dopamine transporter: Mutation of an intracellular tyrosine constitutively alters the conformational equilibrium of the transport cycle. Proc Natl Acad Sci U S A 99:1683–1688

    Article  PubMed  CAS  Google Scholar 

  • McArthur JC, Steiner J, Sacktor N, Nath A (2010) Human immunodeficiency virus-associated neurocognitive disorders: Mind the gap. Ann Neurol 67:699–714

    PubMed  CAS  Google Scholar 

  • Meade CS, Conn NA, Skalski LM, Safren SA (2011a) Neurocognitive impairment and medication adherence in HIV patients with and without cocaine dependence. J Behav Med 34:128–138

    Article  PubMed  Google Scholar 

  • Meade CS, Lowen SB, MacLean RR, Key MD, Lukas SE (2011b) fMRI brain activation during a delay discounting task in HIV-positive adults with and without cocaine dependence. Psychiatry Res 192:167–175

    Article  PubMed  Google Scholar 

  • Midde NM, Gomez AM, Zhu J (2012) HIV-1 Tat protein decreases dopamine transporter cell surface expression and vesicular monoamine transporter-2 function in Rat striatal synaptosomes. J Neuroimmune Pharmacol 7:629–639

    Article  PubMed  Google Scholar 

  • Moritz AE, Foster JD, Gorentla BK, Mazei-Robison MS, Yang JW, Sitte HH, Blakely RD, Vaughan RA (2013) Phosphorylation of dopamine transporter serine 7 modulates cocaine analog binding. J Biol Chem 288:20–32

    Article  PubMed  CAS  Google Scholar 

  • Nath A, Clements JE (2011) Eradication of HIV from the brain: Reasons for pause. AIDS 25:577–580

    Article  PubMed  Google Scholar 

  • Nath A, Maragos WF, Avison MJ, Schmitt FA, Berger JR (2001) Acceleration of HIV dementia with methamphetamine and cocaine. J Neurovirol 7:66–71

    Article  PubMed  CAS  Google Scholar 

  • Norregaard L, Frederiksen D, Nielsen EO, Gether U (1998) Delineation of an endogenous zinc-binding site in the human dopamine transporter. EMBO J 17:4266–4273

    Article  PubMed  CAS  Google Scholar 

  • Peloponese JM Jr, Gregoire C, Opi S, Esquieu D, Sturgis J, Lebrun E, Meurs E, Collette Y, Olive D, Aubertin AM, Witvrow M, Pannecouque C, De Clercq E, Bailly C, Lebreton J, Loret EP (2000) 1H-13C Nuclear magnetic resonance assignment and structural characterization of HIV-1 Tat protein. C R Acad Sci III 323:883–894

    Article  PubMed  CAS  Google Scholar 

  • Purohit V, Rapaka R, Shurtleff D (2011) Drugs of abuse, dopamine, and HIV-associated neurocognitive disorders/HIV-associated dementia. Mol Neurobiol 44:102–110

    Article  PubMed  CAS  Google Scholar 

  • Reith ME, Berfield JL, Wang LC, Ferrer JV, Javitch JA (2001) The uptake inhibitors cocaine and benztropine differentially alter the conformation of the human dopamine transporter. J Biol Chem 276:29012–29018

    Article  PubMed  CAS  Google Scholar 

  • Richfield EK (1993) Zinc modulation of drug binding, cocaine affinity states, and dopamine uptake on the dopamine uptake complex. Mol Pharmacol 43:100–108

    PubMed  CAS  Google Scholar 

  • Robertson KR, Smurzynski M, Parsons TD, Wu K, Bosch RJ, Wu J, McArthur JC, Collier AC, Evans SR, Ellis RJ (2007) The prevalence and incidence of neurocognitive impairment in the HAART era. AIDS 21:1915–1921

    Article  PubMed  Google Scholar 

  • Shan J, Javitch JA, Shi L, Weinstein H (2011) The substrate-driven transition to an inward-facing conformation in the functional mechanism of the dopamine transporter. PLoS One 6:e16350

    Article  PubMed  Google Scholar 

  • Sulzer D, Sonders MS, Poulsen NW, Galli A (2005) Mechanisms of neurotransmitter release by amphetamines: A review. Prog Neurobiol 75:406–433

    Article  PubMed  CAS  Google Scholar 

  • Torres GE, Amara SG (2007) Glutamate and monoamine transporters: new visions of form and function. Curr Opin Neurobiol 17:304–312

    Article  PubMed  CAS  Google Scholar 

  • Tozzi V, Balestra P, Bellagamba R, Corpolongo A, Salvatori MF, Visco-Comandini U, Vlassi C, Giulianelli M, Galgani S, Antinori A, Narciso P (2007) Persistence of neuropsychologic deficits despite long-term highly active antiretroviral therapy in patients with HIV-related neurocognitive impairment: Prevalence and risk factors. J Acquir Immune Defic Syndr 45:174–182

    Article  PubMed  Google Scholar 

  • Wang GJ, Chang L, Volkow ND, Telang F, Logan J, Ernst T, Fowler JS (2004) Decreased brain dopaminergic transporters in HIV-associated dementia patients. Brain 127:2452–2458

    Article  PubMed  Google Scholar 

  • Westendorp MO, Frank R, Ochsenbauer C, Stricker K, Dhein J, Walczak H, Debatin KM, Krammer PH (1995) Sensitization of T cells to CD95-mediated apoptosis by HIV-1 Tat and gp120. Nature 375:497–500

    Article  PubMed  CAS  Google Scholar 

  • Xiao H, Neuveut C, Tiffany HL, Benkirane M, Rich EA, Murphy PM, Jeang KT (2000) Selective CXCR4 antagonism by Tat: Implications for in vivo expansion of coreceptor use by HIV-1. Proc Natl Acad Sci U S A 97:11466–11471

    Article  PubMed  CAS  Google Scholar 

  • Zhao Y, Terry D, Shi L, Weinstein H, Blanchard SC, Javitch JA (2010) Single-molecule dynamics of gating in a neurotransmitter transporter homologue. Nature 465:188–193

    Article  PubMed  CAS  Google Scholar 

  • Zhu J, Apparsundaram S, Bardo MT, Dwoskin LP (2005) Environmental enrichment decreases cell surface expression of the dopamine transporter in rat medial prefrontal cortex. J Neurochem 93:1434–1443

    Article  PubMed  CAS  Google Scholar 

  • Zhu J, Mactutus CF, Wallace DR, Booze RM (2009) HIV-1 Tat protein-induced rapid and reversible decrease in [3H]dopamine uptake: Dissociation of [3H]dopamine uptake and [3H]2beta-carbomethoxy-3-beta-(4-fluorophenyl)tropane (WIN 35,428) binding in rat striatal synaptosomes. J Pharmacol Exp Ther 329:1071–1083

    Article  PubMed  CAS  Google Scholar 

  • Zhu J, Ananthan S, Mactutus CF, Booze RM (2011) Recombinant human immunodeficiency virus-1 transactivator of transcription1-86 allosterically modulates dopamine transporter activity. Synapse 65:1251–1254

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This research was supported by grants from the National Institutes of Health to Jun Zhu (DA024275, DA026721), Chang-Guo Zhan (DA032910, DA035552), and Rosemarie Booze (DA013137, HD043680).

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The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Department of Drug Discovery and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, 715 Sumter Street, Columbia, SC, 29208, USA

    Narasimha M. Midde, Adrian M. Gomez & Jun Zhu

  2. Department of Psychology, University of South Carolina, Columbia, SC, 29208, USA

    Rosemarie M. Booze & Jun Zhu

  3. Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY, 40536, USA

    Xiaoqin Huang & Chang-Guo Zhan

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  1. Narasimha M. Midde
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  2. Xiaoqin Huang
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  5. Chang-Guo Zhan
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  6. Jun Zhu
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Correspondence to Jun Zhu.

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Midde, N.M., Huang, X., Gomez, A.M. et al. Mutation of Tyrosine 470 of Human Dopamine Transporter is Critical for HIV-1 Tat-Induced Inhibition of Dopamine Transport and Transporter Conformational Transitions. J Neuroimmune Pharmacol 8, 975–987 (2013). https://doi.org/10.1007/s11481-013-9464-6

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