Abstract
The TaqIA single-nucleotide polymorphism (SNP), which is the most widely studied genetic polymorphism in addictions, is located at the gene that encodes the RIP kinase ANKK1 near the gene for dopamine receptor D2. The TaqIA SNP is in strong linkage disequilibrium with the SNP rs7118900, which changes the alanine at position 239 to threonine in the ANKK1 protein (Ala239/A2; Thr239/A1). In silico analysis has predicted that this polymorphic substitution creates an additional phosphorylation site in the kinase domain of ANKK1. To investigate the contribution of ANKK1 to the pathophysiology of TaqIA-associated phenotypes, we analyzed transfected HEK293T cells with the human ANKK1-kinaseAla239 and ANKK1-kinaseThr239 variants tagged with GFP. We observed that the ANKK1-kinase is located in both the nucleus and the cytoplasm, suggesting that there is nucleocytoplasmic shuttling of this putative signal transducer. In addition, we found that the Ala239Thr ANKK1-kinase polymorphism exhibited strong expression differences in both the nucleus and the cytoplasm at basal level and when stimulated with the dopamine agonist apomorphine. Specifically, the ANKK1-kinaseThr239 variant showed the highest level of basal protein expression, while ANKK1-kinaseAla239 was 0.64-fold lower. After treatment with apomorphine, ANKK1-kinaseAla239 showed a 2.4-fold increment in protein levels, whereas a 0.67-fold reduction was observed in ANKK1-kinaseThr239. Thus, here we provide the first evidence of functional ANKK1 differences that are marked by TaqIA and could be associated with vulnerability to addiction.
Similar content being viewed by others
References
Birbach A, Gold P, Binder BR, Hofer E, de Martin R, Schmid JA (2002) Signaling molecules of the NF-kappa B pathway shuttle constitutively between cytoplasm and nucleus. J Biol Chem 277:10842–10851
Blum K, Noble EP, Sheridan PJ, Montgomery A, Ritchie T, Jagadeeswaran P, Nogami H, Briggs AH, Cohn JB (1990) Allelic association of human dopamine D2 receptor gene in alcoholism. JAMA 263:2055–2060
Bogerd HP, Fridell RA, Benson RE, Hua J, Cullen BR (1996) Protein sequence requirements for function of the human T-cell leukemia virus type 1 Rex nuclear export signal delineated by a novel in vivo randomization-selection assay. Mol Cell Biol 16:4207–4214
David SP, Strong DR, Munafo MR, Brown RA, Lloyd-Richardson EE, Wileyto PE, Evins EA, Shields PG, Lerman C, Niaura R (2007) Bupropion efficacy for smoking cessation is influenced by the DRD2 Taq1A polymorphism: analysis of pooled data from two clinical trials. Nicotine Tob Res 9:1251–1257
Dick DM, Wang JC, Plunkett J, Aliev F, Hinrichs A, Bertelsen S, Budde JP, Goldstein EL, Kaplan D, Edenberg HJ, Nurnberger J Jr, Hesselbrock V, Schuckit M, Kuperman S, Tischfield J, Porjesz B, Begleiter H, Bierut LJ, Goate A (2007) Family-based association analyses of alcohol dependence phenotypes across DRD2 and neighboring gene ANKK1. Alcohol Clin Exp Res 31:1645–1653
Gelernter J, Yu Y, Weiss R, Brady K, Panhuysen C, Yang BZ, Kranzler HR, Farrer L (2006) Haplotype spanning TTC12 and ANKK1, flanked by the DRD2 and NCAM1 loci, is strongly associated to nicotine dependence in two distinct American populations. Hum Mol Genet 15:3498–3507
Hoenicka J, Quinones-Lombrana A, Espana-Serrano L, Alvira-Botero X, Kremer L, Perez-Gonzalez R, Rodriguez-Jimenez R, Jimenez-Arriero MA, Ponce G, Palomo T (2010) The ANKK1 gene associated with addictions is expressed in astroglial cells and upregulated by apomorphine. Biol Psychiatry 67:3–11
Horton P, Park KJ, Obayashi T, Fujita N, Harada H, Adams-Collier CJ, Nakai K (2007) WoLF PSORT: protein localization predictor. Nucleic Acids Res 35:W585–W587
la Cour T, Kiemer L, Molgaard A, Gupta R, Skriver K, Brunak S (2004) Analysis and prediction of leucine-rich nuclear export signals. Protein Eng Des Sel 17:527–536
Laakso A, Pohjalainen T, Bergman J, Kajander J, Haaparanta M, Solin O, Syvalahti E, Hietala J (2005) The A1 allele of the human D2 dopamine receptor gene is associated with increased activity of striatal L-amino acid decarboxylase in healthy subjects. Pharmacogenet Genomics 15:387–391
Lucht M, Samochowiec A, Samochowiec J, Jasiewicz A, Grabe HJ, Geissler I, Rimmbach C, Rosskopf D, Grzywacz A, Wysiecka JP, Tybura P, Brzuchalski B, Bienkowski P (2010) Influence of DRD2 and ANKK1 genotypes on apomorphine-induced growth hormone (GH) response in alcohol-dependent patients. Prog Neuropsychopharmacol Biol Psychiatry 34:45–49
Meylan E, Tschopp J (2005) The RIP kinases: crucial integrators of cellular stress. Trends Biochem Sci 30:151–159
Meylan E, Martinon F, Thome M, Gschwendt M, Tschopp J (2002) RIP4 (DIK/PKK), a novel member of the RIP kinase family, activates NF-kappa B and is processed during apoptosis. EMBO Rep 3:1201–1208
Munafo MR, Matheson IJ, Flint J (2007) Association of the DRD2 gene Taq1A polymorphism and alcoholism: a meta-analysis of case-control studies and evidence of publication bias. Mol Psychiatry 12:454–461
Munafo MR, Timpson NJ, David SP, Ebrahim S, Lawlor DA (2009) Association of the DRD2 gene Taq1A polymorphism and smoking behavior: a meta-analysis and new data. Nicotine Tob Res 11:64–76
Neville MJ, Johnstone EC, Walton RT (2004) Identification and characterization of ANKK1: a novel kinase gene closely linked to DRD2 on chromosome band 11q23.1. Hum Mutat 23:540–545
Noble EP (2003) D2 dopamine receptor gene in psychiatric and neurologic disorders and its phenotypes. Am J Med Genet B 116:103–125
Ponce G, Hoenicka J, Jimenez-Arriero MA, Rodriguez-Jimenez R, Aragues M, Martin-Sune N, Huertas E, Palomo T (2008) DRD2 and ANKK1 genotype in alcohol-dependent patients with psychopathic traits: association and interaction study. Br J Psychiatry 193:121–125
Ponce G, Perez-Gonzalez R, Aragues M, Palomo T, Rodriguez-Jimenez R, Jimenez-Arriero MA, Hoenicka J (2009) The ANKK1 kinase gene and psychiatric disorders. Neurotox Res 16:50–59
Ramnarain DB, Paulmurugan R, Park S, Mickey BE, Asaithamby A, Saha D, Kelliher MA, Mukhopadhyay P, Banani F, Madden CJ, Wright PS, Chakravarty S, Habib AA (2008) RIP1 links inflammatory and growth factor signaling pathways by regulating expression of the EGFR. Cell Death Differ 15:344–353
Samochowiec J, Ladehoff M, Pelz J, Smolka M, Schmidt LG, Rommelspacher H, Finckh U (2000) Predominant influence of the 3′-region of dopamine D2 receptor gene (DRD2) on the clinical phenotype in German alcoholics. Pharmacogenetics 10:471–475
Schmidt LG, Dettling M, Graef KJ, Heinz A, Kuhn S, Podschus J, Rommelspacher H (1996) Reduced dopaminergic function in alcoholics is related to severe dependence. Biol Psychiatry 39:193–198
Shen YC, Chen SF, Chen CH, Lin CC, Chen SJ, Chen YJ, Luu SU (2009) Effects of DRD2/ANKK1 gene variations and clinical factors on aripiprazole efficacy in schizophrenic patients. J Psychiatr Res 43:600–606
Yang Y, Ma J, Chen Y, Wu M (2004) Nucleocytoplasmic shuttling of receptor-interacting protein 3 (RIP3): identification of novel nuclear export and import signals in RIP3. J Biol Chem 279:38820–38829
Yang BZ, Kranzler HR, Zhao H, Gruen JR, Luo X, Gelernter J (2007) Association of haplotypic variants in DRD2, ANKK1, TTC12 and NCAM1 to alcohol dependence in independent case control and family samples. Hum Mol Genet 16:2844–2853
Zhang Y, Bertolino A, Fazio L, Blasi G, Rampino A, Romano R, Lee ML, Xiao T, Papp A, Wang D, Sadee W (2007) Polymorphisms in human dopamine D2 receptor gene affect gene expression, splicing, and neuronal activity during working memory. Proc Natl Acad Sci USA 104:20552–20557
Acknowledgments
This study was supported by the Fondo de Investigación Sanitaria, Instituto Salud Carlos III, Grant No. 08/0529. CIBERSAM is an initiative of the Instituto de Salud Carlos III.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Garrido, E., Palomo, T., Ponce, G. et al. The ANKK1 Protein Associated with Addictions has Nuclear and Cytoplasmic Localization and Shows a Differential Response of Ala239Thr to Apomorphine. Neurotox Res 20, 32–39 (2011). https://doi.org/10.1007/s12640-010-9219-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12640-010-9219-6