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Characterization of intracellular aggregates using fluorescently-tagged polyglutamine-expanded androgen receptor

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Abstract

Spinal bulbar muscular atrophy (SBMA) is a classic CAG-repeat neurodegenerative disease. It is caused by expansion of a polyglutamine (polyGln) tract in the androgen receptor (AR). Recent evidence has indicated a potential role for nuclear and cytoplasmic inclusions in the pathogenesis of these diseases. We have used blue and green fluorescently-tagged AR to show that both wild-type (WT) and poly-Gln-expanded full-length AR can form aggregates and that aggregation is not related to cytotoxicity. Twenty to thirty-five percent of all cell types transfected into COS cells showed aggregation containing both amino-and carboxy-terminal fluorescent tags. The aggregates reacted with (F39.4.1), an anti-AR antibody and with 1C2, an expanded polyGln tract antibody. Western analysis of protein extracts revealed little evidence of proteolysis although some cleavage of the fusion proteins was seen. The general caspase inhibitor, Z-DEVD-FMK, did not affect aggregation in either wild type or polyGln-expanded GFP-AR transfected cells. Surprisingly, addition of Mibolerone a synthetic androgen significantly decrease inclusion formation in both WT and polyGln-expanded AR-transfected cells. Overall, we show that both WT and polyGln expanded full-length AR are found in aggregates and that proteolysis is not a requirement for aggregation. Our results also suggest that toxicity is not related to intracellular aggregation of polyGln expanded AR.

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References

  • Abdullah, A.A.R., Trifiro, M.A., Panet-Raymond, V., Alvar-ado, C, de Tourreil, S., Frankel, D., Schipper, H.M. and Pinsky, L. (1998) Spinobulbar muscular atrophy: poly-Gln-expanded androgen receptor is proteolytically resistantin vitro and processed abnormally in trans-fected cells. Hum. Mol. Genet.7, 379–384.

    Article  PubMed  CAS  Google Scholar 

  • Becher, M.W., Kotzuk, J.A., Sharp, A.H., Davies, S.W., Bates, G.P., Price, D.L. and Ross, C.A. (1998) Intranuclear neuronal inclusions in Huntington’s disease and dentatoru-bral and pallidoluysian atrophy: correlation between the density of inclusions and IT15 CAG triplet repeat length. Neurobiol. Dis.4, 387–397.

    Article  PubMed  CAS  Google Scholar 

  • Beitel, L.K., Prior, L., Vasiliou, D.M., Gottlieb, B., Kaufman, M., Lumbroso, R., Alvarado, C, McGillivray, B., Trifiro, M. and Pinsky, L. (1994) Complete androgen insensitiv-ity due to mutations in the probable a-helical segments of the DNA-binding domain in the human androgen receptor. Hum. Mol. Genet.3, 21–27.

    Article  PubMed  CAS  Google Scholar 

  • Bok, K.S., Rhim, H., Yoo, Y.D., Choi, E., Ahn, K., Kim, I.H. and Kang, S. (1999) Expanded polyglutamine tract itself induces cell death in cultured cells. Molecules and Cells9, 398–402.

    PubMed  CAS  Google Scholar 

  • Brooks, B.P., Paulson, H.L., Merry, D.E., Salazar-Grueso, E.F., Brinkman, A.O., Wilson, E.M., Fischbeck, K.H. (1997) Characterization of an expanded glutamine repeat androgen receptor in a neuronal cell culture system. Neurobiol. Dis.4, 313–323.

    Article  Google Scholar 

  • Cooper, J.K., Schilling, G., Peters, M.F., Herring, W.J., Sharp, A.H., Kaminsky, Z., Masone, J., Khan, F.A., Delanoy, M., Borchelt, D.R., Dawson, V.L., Dawson, T.M. and Ross, C.A. (1998) Truncated N-terminal fragments of hunting-tin with expanded glutamine repeats form nuclear and cytoplasmic aggregates in cell culture. Hum. Mol. Genet.7, 783–790.

    Article  PubMed  CAS  Google Scholar 

  • Cummings, C.J., Mancini, M.A., Antalffy, B., DeFranco, D.B., Orr, H.T. and Zoghbi, H.Y. (1998) Chaperone suppression of aggregation and altered subcellular proteasome localization imply protein misfolding in SCA1. Nat. Genet.19, 148–154.

    Article  PubMed  CAS  Google Scholar 

  • Cummings, C.J. and Zoghbi, H.Y. (2000) Fourteen and counting: unraveling trinucleotide repeat diseases. Hum. Mol. Genet.9, 909–916.

    Article  PubMed  CAS  Google Scholar 

  • David, G., Abbas, N., Stevanin, G., Durr, A., Yvert, G., Cancel, G., Weber, C, Imbert, G., Saudou, F., Antoniou, E. et al. (1997) Cloning of the SCA7 gene reveals a highly unstable CAG repeat expansion. Nat. Genet.17, 65–70.

    Article  PubMed  CAS  Google Scholar 

  • Davies, S.W., Turmaine, M., Cozens, B.A., DiFiglia, M., Sharp, A.H., Ross, C.A., Scherzinger, E., Wanker, E.E., Mangiarini, L. and Bates, G.P. (1997) Formation of neuronal intranuclear inclusions underlies the neurology dysfunction in mice transgenic for the HD mutation. Cell90, 537–548.

    Article  PubMed  CAS  Google Scholar 

  • Davies, S.W., Beardsall, K., Turmaine, M., DiFiglia, M., Aronin, N. and Bates, G.P. (1998) Are neuronal intranuclear inclusions the common neuropathology of triplet-repeat disorders with polyglutamine-repeat expansions? Lancet351, 131–133.

    Article  PubMed  CAS  Google Scholar 

  • DiFiglia, M., Sapp, E., Chase, K.O., Davies, S.W., Bates, G.P., Vonsattel, J.P. and Aronin, N. (1997) Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science277, 1990–1993.

    Article  PubMed  CAS  Google Scholar 

  • Ellerby, L.M., Hackam, A.S., Propp, S.S., Ellerby, H.M., Rabi-zadeh, S., Gashman, N.R., Trifiro, M.A., Pinsky, L., Wellington, C.L., Salvesen, G.S., Hayden, M.R. and Bredesen, D.E. (1999) Kennedy’s disease: caspase cleavage of the androgen receptor is a crucial event in cytotoxicity. J. Neurochem.72, 185–195.

    Article  PubMed  CAS  Google Scholar 

  • Georget, V., Lobaccaro, J.M., Terouanne, B., Mangeat, P., Nicolas, J.-C. and Sultan, C. (1997) Trafficking of the androgen receptor in living cells with fused green fluorescent protein-androgen receptor. Mol. Cell. Endocrinol.129, 17–26.

    Article  PubMed  CAS  Google Scholar 

  • Goldberg, Y.Pl, Nicholson, D.W., Rasper, D.M., Kalchman, M.A., Koide, H.B., Graham, R.K., Bromm, M., Kazemi-Esfarjani, P., Thornberry, N. A., Vaillancourt, J.P. and Hayden, M.R. (1996) Cleavage of huntingtin by apopain, a proapoptotic cysteine protease is modulated by the polyglutamine tract. Nat. Genet.13, 442–449.

    Article  PubMed  CAS  Google Scholar 

  • Gourfinkel-An, I., Cancel, G., Duyckaerts, C, Faucheux, B., Hauw, J.J., Trottier, Y., Brice, A., Agid, Y. and Hirsch, E.C. (1998) Neuronal distribution of intranuclear inclusions in Huntington’s disease with adult onset. Neurore-port9, 1823–1826.

    CAS  Google Scholar 

  • Gutekunst, C-A., Li, S-H., Yi, H., Mulroy, J.S., Kuemmerle, S., Jones, R., Rye, D., Ferrante, R.J., Hersch, S.M. and Li, X-P. (1999) Nuclear and neutropil aggregates in Huntington’s disease: relationship to neuropathology. J. Neu-rosci.19, 2522–2534.

    CAS  Google Scholar 

  • Hackam, A., Singaraja, T., Wellington, C.L., Metzler, M., McCutcheon, K., Zhang, T., Kalchaman, M. and Hayden, M.R. (1998) The influence of huntingtin protein size on nuclear localization and cellular toxicity. J. Cell Biol.141, 1097–1105.

    Article  PubMed  CAS  Google Scholar 

  • Hodgson, J.G., Agopyan, N., Gutenunst, C-A., Leavitt, B.R., LePlane, F., Singaraja, R., Smith, D.J., Bissada, N., McCutcheon, K., Nasir, J., Jamot, L., Li, X-J., Stevens, M.E., Rosemond, E., Roder, J.C., Phillips, A.G., Rubin, E.M., Hersch, S.M. and Hayden, M.R. (1999) A YAC mouse model for Huntington’s Disease with full-length mutant huntington, cytoplasmic toxicity and selective striatal neurodegeneration. Neuron23:181–192.

    Article  PubMed  CAS  Google Scholar 

  • Holmberg, M., Duyckaerts, C, Durr, A., Cancel, G., Gourfinkel-An, I., Damier, P., Faucheux, B., Trottier, Y., Hirsch, E.C, Agid, Y. and Brice, A. (1998) Spinocerebellar ataxia type 7 (SCA7): a neurodegenerative disorder with neuronal intranuclear inclusions. Hum. Mol. Genet.7, 913–918.

    Article  PubMed  CAS  Google Scholar 

  • Huntington’s Disease Collaborative Research Group (1993) A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell72, 971–983.

    Article  Google Scholar 

  • Igarashi, S., Koide, R., Shimohata, T., Yamada, M., Hayashi, Y., Takano, H., Date, H., Oyake, M., Sato, T., Sato, A. et al. (1998) Suppression of aggregate formation and apop-tosis by transglutaminase inhibitors in cells expressing truncated DRPLA protein with an expanded polyglutamine stretch. Nat. Genet.18, 111–117.

    Article  PubMed  CAS  Google Scholar 

  • Ikeda, H., Masahiro, Y., Satoshi, S., Yoshiya, A., Shuh, N. and Kakizuka, A. (1996) Expanded polyglutamine in Mach-ado-Joseph disease protein induces cell deathin vitro andin vivo. Nat. Genet.13, 196–202.

    CAS  Google Scholar 

  • Imbert, G., Saudou, F., Yvert, G., Devys, D., Trottier, Y., Gar-nier, J.M., Weber, C, Mandel, J.L., Cancel, G., Abbas, N. et al. (1996) Cloning of the gene for spinocerebellar ataxia 2 reveals a locus with high sensitivity to expanded CAG/glutamine repeats. Nat. Genet.14, 285–291.

    Article  PubMed  CAS  Google Scholar 

  • Jenster, G., van der Korput, J.A. G. M., van Vroonhoven, C.C. J., van der Kwast. T.H., Trapman, J. and Brinkmann, A.O., (1991) Domains of the human androgen receptor involved in steroid binding, transcriptional activation and subcellular localization. Mol Endocrinol5, 1394–1404.

    Article  Google Scholar 

  • Kahlem, P., Terre, C, Green, H. and Djian, P. (1996) Peptides containing glutamine repeats as substrates for trans-glutaminase-catalyzed cross-linking: relevance to diseases of the nervous system. Proc. Natl. Acad. Sci. USA93, 14580–14585.

    Article  PubMed  CAS  Google Scholar 

  • Kawaguchi, Y., Okamoto, T., Taniwaki, M., Aizawa, M., Inoue, M., Katayama, S., Kawakami, H., Nakamura, S., Nishimura, M., Akiguchi, I. et al. (1994) CAG expansions in a novel gene for Machado-Joseph disease at chromosome 14q32.1. Nat. Genet.8, 221–222.

    Article  PubMed  CAS  Google Scholar 

  • Kazemi-Esfarjani, P., Trifiro, M. and Pinsky, L. (1995) Evidence for a repressive of the long polyglutamine tract in the human androgen receptor: possible pathogenetic relevance for (CAG)n-expanded neuronopathies. Hum. Mol. Genet.4, 523–527.

    Article  PubMed  CAS  Google Scholar 

  • Kazemi-Esfarjani, P. and Benzer, S. (2000) Genetic suppression of polyglutamine toxicity inDrosophila. Science287, 1837–1840.

    Article  CAS  Google Scholar 

  • Kazantsev, A., Preisinger E., Dranovsky, A., Goldgaber, D. and Housman, D. (1999) Insoluble detergent-resistant aggregates form between pathological and nonpatholog-ical lengths of polyglutamine in mammalian cells. Proc. Natl. Acad. Sci. USA96, 11404–11409.

    Article  PubMed  CAS  Google Scholar 

  • Kennedy, W.R., Alter, M. and Sung, J.H. (1968) Progressive proximal spinal and bulbar muscular atrophy of late onset. A sex-linked trait. Neurology18, 671–680.

    CAS  Google Scholar 

  • Kim, M., Lee, H.-S., LaForet, G., Mclntyre, C, Martin, E.J., Chang, P., Kim, T.W., Williams, M., Reddy, P.H., Tagle, D., Boyce, F.M., Won, L., Heller, A., Aronin, N. and DiFiglia, M. (1999) Mutant huntingtin expression in clonal striatal cells: dissociation of inclusion formation and neuronal survival by caspase inhibition. J. Neurosci.19, 964–973.

    PubMed  CAS  Google Scholar 

  • Kim, T.-W. and Tanzi, R.E. (1998) Neuronal intranuclear inclusions in polyglutamine diseases: nuclear weapons or nuclear fallout? Neuron21, 657–659.

    Article  PubMed  CAS  Google Scholar 

  • Klement, I.A., Skiner, P.J., Kaytor, M.D., Yi, H., Hersch, S.M., Clark, H.B., Zoghbi, H.Y. and Orr, H.T. (1998) Ataxin-1 nuclear localization and aggregation: role in poly-glutamine-induced disease inSCA1 transgenic mice. Cell95, 41–53.

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi, Y., Miwa, S., Merry, D.E., Kume, A., Mei, L., Doyu, M. and Sobue, G. (1998) Caspase-3 cleaves the expanded androgen receptor protein of spinal and bulbar muscular atrophy in a polyglutamine repeat length-dependent manner. Biochem. Biophys. Res. Com-mun.252, 145–150.

    Article  CAS  Google Scholar 

  • Koide, R., Ikeuchi, T., Onodera, O., Tanaka, H., Igarashi, S., Endo, K., Takahashi, H., Kondo, R., Ishikawa, A., Hay-ashi, T. et al. (1994) Unstable expansion of CAG repeat in hereditary dentatorubral-pallidoluysian atrophy (DRPLA). Nat. Genet.6, 9–13.

    Article  PubMed  CAS  Google Scholar 

  • Koob, M.D., Benzow, K.A., Bird, T.D., Day, J.W., Moseley, M.L. and Ranum, L.P. (1998) Rapid cloning of expanded trinucleotide repeat sequences from genomic DNA. Nat. Genet.18, 72–75.

    Article  PubMed  CAS  Google Scholar 

  • Koyano, S., Uchihara, T., Fujigasaki, H., Nakamra, A., Yagis-hita, S. and Iwabuchi, K. (1999) Neuronal intranuclear inclusions in spinocereballar ataxia type 2: triple-labeling immunofluorescent study. Neuroscience Letters273, 117–120.

    Article  PubMed  CAS  Google Scholar 

  • Kuida, K., Zheng, T.S., Na, S., Kuan, C, Yang, D., Kara-suyama, H., Rakic, P. and Flavell, R.A. (1996) Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature384, 368–372.

    Article  PubMed  CAS  Google Scholar 

  • Kummerle, S., Gutekunst, C-A., Klein, A.M., Li, X-J., Li, S-H., Beal, M.F., Hersch, S.M. And Ferrante, R.J. (1999) Huntington aggregates may not predict neuronal death in Huntington’s Disease. Ann Neurol.46: 842–849.

    Article  Google Scholar 

  • LaSpada, A.R., Wilson, E.M., Lubahn, D.B., Harding, A.E. and Fischbeck, K.H. (1991) Androgen receptor gene mutations in X-linked spinal and bulbar muscular atrophy. Nature352, 77–79.

    Article  CAS  Google Scholar 

  • Lescure, A., Lutz, Y., Eberhard, D., Jacq, X., Krol, A., Grummt, I., Davidson, I., Chambon, P. and Tora, L. (1994) The N-terminal domain of the human TATA-binding protein plays a role in transcription from TATA-containing RNA polymerase II and III promoters. EMBO J.13, 1166–1175.

    PubMed  CAS  Google Scholar 

  • Li, M., Miwa, S., Yasushi, K., Merry, D.E., Yamamoto, M., Tanaka, F., Doyu, M., Hashizume, Y., Fishbeck K.H. and Sobue, G. (1998a) Nuclear inclusions of the androgen receptor protein in spinal and bulbar muscular atrophy. Ann Neurol.44, 249–254.

    Article  PubMed  CAS  Google Scholar 

  • Li, M., Nakagomi, Y., Kobayashi, Y., Merry, D.E., Tanaka, F., Doyu, M., Mitsuma, T., Hashizume, Y., Fischbeck, K.H. and Sobue, G. (1998b) Nonneural nuclear inclusions of androgen receptor protein in spinal and bulbar muscular atrophy. Am. J. Pathol.153, 695–701.

    PubMed  CAS  Google Scholar 

  • Lunkes, A. and Mandel, J.-L. (1997) Polyglutamines, nuclear inclusions and neurodegeneration. Nat. Med.3, 1201–1202.

    Article  PubMed  CAS  Google Scholar 

  • Martindale, D., Hackam, A., Wieczorek, A., Ellerby, L., Wellington, C, McCutcheon, K., Singaraja, R., Kazemi-Esfar-jani, P., Devon, R., Kim, S.U., Bredesen, D., Tufaro, F. and Hayden, M.R. (1998) Length of huntingtin and its polyglutamine tract influences localization and frequency of intracellular aggregates. Nat. Genet.18, 150–154.

    Article  PubMed  CAS  Google Scholar 

  • Matilla, A., Roberson, E.D., Banfi, S., Morales, J., Armstrong, D.L., Burright, E.N., Orr, H.T., Sweatt, J.D., Zoghbi, H.Y. and Matzuk, M.M. (1998) Mice lacking ataxin-1 display learning deficits and decreased hippocampal paired-pulse facilitation. J. Neurosci.18, 5508–5516.

    PubMed  CAS  Google Scholar 

  • Merry D.E., Kobayashi, Y., Bailey C.K., Taye, A.A., Fischbeck, K.H. (1998) Cleavage, aggregation and toxicity of the expanded androgen receptor in spinal and bulbar muscular atrophy. Hum. Mol. Genet.7: 693–701.

    Article  PubMed  CAS  Google Scholar 

  • Miyashita, T., Nagao, K., Ohmi, K., Yanagisawa, H., Oka-mura-Oho, Y. and Yamada, M. (1998) Intracellular aggregate formation of dentatorubral-pallidoluysian atrophy (DRPLA) protein with the extended polyglutamine. Biochem. Biophys. Res. Commun.249, 96–102.

    Article  PubMed  CAS  Google Scholar 

  • Miyashita, T., Okamura-Oho, Y., Mito, Y., Nagafuchi, S. and Yamada, M. (1997) Dentatorubral pallidoluysian atrophy (DRPLA) protein is cleaved by caspase-3 during apoptosis. J. Biol. Chem.272, 29238–29242.

    Article  PubMed  CAS  Google Scholar 

  • Miwa, K., Asano, M., Horai, R., Iwakura, Y., Nagata, S. and Suda, T. (1998) Caspase 1-independent IL-1 beta release and inflammation induced by the apoptosis induced Fas ligand. Nat. Med.4, 1287–1292.

    Article  PubMed  CAS  Google Scholar 

  • Moulder, K.L., Onodera, O., Burke, J.R., Strittmatter, W.J. and Johnson E.M. Jr. (1999) Generation of neuronal intranuclear inclusions by polyglutamine-GFP: analysis of inclusion clearance and toxicity as a function of polyglutamine length. J. Neurosci.9, 705–715.

    Google Scholar 

  • Nagafuchi, S., Yansgisawa, H., Sato, K., Shirayama, T., Ohsaki, E., Bundo, M., Takeda, T., Tadokoro, K., Kendo, I., Murayama, N. et al. (1994) Denatorubal and pallidoluysian atrophy expansion of an unstable CAG trinucleotide on chromosome 12p. Nature Genet.6, 14–18.

    Article  PubMed  CAS  Google Scholar 

  • Okajima, T., Tanabe, T. and Yasuda, T. (1993) Nonurea sodium dodecyl sulfate-polyacrylamide gel electrophoresis with high-molarity buffers for the separation of proteins and peptides. Anal. Biochem.211, 293–300.

    Article  PubMed  CAS  Google Scholar 

  • Ordway, J.M., Tallaksen-Greene, S., Gutekunst, C.A., Bernstein, E.M., Cearley, J.A., Wiener, H.W., Dure, L.S. 4th., Lindsey, R., Hersch, S.M., Jope, R.S., Albin, R.L. and Det-loff, P.J. (1997) Ectopically expressed CAG repeats cause intranuclear inclusions and a progressive late onset neurological phenotype in the mouse. Cell91, 753–763.

    Article  PubMed  CAS  Google Scholar 

  • Orr, H.T., Chung, M.Y., Banfi, S., Kwiatkowski, T.J., Serva-dio, A., Beaudet, A.L., McCall, A.E., Duvick, L.A., Ranum, L.P. and Zoghbi, H.Y. (1993) Expansion of an unstable trinucleotide CAG repeat in spinocerebellar ataxia type 1. Nat. Genet.4, 221–226.

    Article  PubMed  CAS  Google Scholar 

  • Paulson, H.L., Perez, M.K., Trottier, Y., Trojanowski, J.Q., Subramony, S.H., Das, S.S., Vig, P., Mandel, J.-L., Fisch-beck, K.H. and Pittman, R.N. (1997) Intranuclear inclusions of expanded polyglutamine protein in spinocerebellar ataxia type 3. Neuron19, 333–344.

    Article  PubMed  CAS  Google Scholar 

  • Paulson, H.L. (1999) Protein fate in neurodegenerative pro-teinopathies: poIyGln diseases join the (mis)fold. Am J Hum Genet64, 339–345.

    Article  PubMed  CAS  Google Scholar 

  • Perez, M.K., Paulson, H.L., Pendse, S.J., Saionz, S., Bonini, N.M. and Pittman, R.N. (1998) Recruitment and the role of nuclear localization in polyglutamine-mediated agre-gation. J. Cell Biol.143, 1457–1470.

    Article  PubMed  CAS  Google Scholar 

  • Perutz, M.F., Johnson, T., Suzuki, M. and Finch, J.T. (1994) Glutamine repeats as polar zippers: their possible role in inherited neurodegenerative diseases. Proc. Natl. Acad. Sci. USA91, 5355–5358.

    Article  PubMed  CAS  Google Scholar 

  • Pulst, S.M., Nechiporuk, A., Nechiporuk, T., Gispert, S., Chen, X.N., Lopes-Cendes, I., Pearlman, S., Starkman, S., Orozco-Diaz, G., Lunkes, A. et al. (1996) Moderate expansion of a normally biallelic trinucleotide repeat in spinocerebellar ataxia type 2. Nat. Genet.14, 269–276.

    Article  PubMed  CAS  Google Scholar 

  • Reddy, P.H., Vinod, C, Williams, M, Miller G., Whetsell, W.O. and Tagle, D.A. (1999) Transgenic mice expressing mutated full-length HD cDNA: a paradigm for locomotor changes and selective neuronal loss in Huntington’s disease. Phil. Trans. R. Soc. Lond. B.354, 1035–1045.

    Article  CAS  Google Scholar 

  • Ross, C.A. (1997) Intranuclear neuronal inclusions: a common pathogenic mechanism for glutamine-repeat neurodegenerative diseases? Neuron19, 1147–1150.

    Article  PubMed  CAS  Google Scholar 

  • Sanchez, I., Xu, C.-J., Juo, P., Kakizaka, A., Blenis, J. and Yuan, J. (1999) Caspase-8 is required for cell death induced by expanded polyglutamine repeats. Neuron22, 623–633.

    Article  PubMed  CAS  Google Scholar 

  • Sanpei, K., Takano, H., Igarashi, S., Sato, T., Oyake, M., Sasaki, H., Wakisaka, A., Tashiro, K., Ishida, Y., Ikeuchi, T. et al. (1996) Identification of the spinocerebellar ataxia type 2 gene using a direct identification of repeat expansion and cloning technique, DIRECT. Nat. Genet.14, 277–284.

    Article  PubMed  CAS  Google Scholar 

  • Saudou, F., Finkbeiner, S., Devys, D. and Greenberg, M.E. (1998) Huntingtin acts in the nucleus to induce apopto-sis but death does not correlate with the formation of intranuclear inclusions. Cell95, 55–66.

    Article  PubMed  CAS  Google Scholar 

  • Simeoni, S., Mancini, M.A., Stenoien, D.L., Marcelli, M., Weigel, NX., Zanisi, L. and Poletti, A. (2000) Motorneural cell death is not correlated with aggregate formation of androgen receptors containing an elongated polyGln tract. Hum. Mol. Genet.9, 133–144.

    Article  PubMed  CAS  Google Scholar 

  • Sisodia, S.S. (1998) Nuclear inclusions in glutamine repeat disorders: are they pernicious, coincidental, or beneficial? Cell95, 1–4.

    Article  PubMed  CAS  Google Scholar 

  • Skinner, P.J., Koshy, B., Cummings, C.J., Klement, I.A., Helin, K., Servadio, A., Zoghbi, H.Y. and Orr, H.T. (1997) SCA1 pathogenesis involves alterations in nuclear matrix associated structures. Nature389, 971–974.

    Article  PubMed  CAS  Google Scholar 

  • Sobue, G., Hashizume, Y., Mukai, E., Hirayama, M., Mit-suma, T. and Takahashi, A. (1989) X-Iinked recessive bulbospinal neuronopathy. A clinicopathological study. Brain112, 209–232.

    Google Scholar 

  • Stenoien, D.L., Cummings, C.J., Adams, H.P., Mancini, M.G., Patel, K., DeMartino, G.N., Marcelli, M., Weigel, N.L. and Mancini, M.A. (1999) Polyglutamine-expanded androgen receptors form aggregates that sequester heat shock proteins, proteasome components and SRC-1 and are suppressed by the HDJ-2 chaperone. Hum. Mol. Genet.8, 731–741.

    Article  PubMed  CAS  Google Scholar 

  • Trottier, Y., Lutz, Y., Stevanin, G., Imbert, G., Devys, D., Cancel, G., Saudou, F., Weber, C, David, G., Tora, L. et al. (1995) Polyglutamine expansion as a pathological epitope in Huntington’s disease and four dominant cerebellar ataxias. Nature378, 403–406.

    Article  PubMed  CAS  Google Scholar 

  • Warrick, J.M., Paulson, H.L., Gray-Board, G.L., Bui, Q.T., Fis-chbeck, K.H., Pittman, R.N. and Bonini, N.M. (1998) Expanded polyglutamine protein forms nuclear inclusions and causes neural degeneration inDrosophila. Cell93, 939–949.

    Article  PubMed  CAS  Google Scholar 

  • Wellington, C.L., Ellerby, L.M., Hackam, A.S., Margolis, R.L., Trifiro, M.A., Singaraja, R., McCutcheon, K., Salvesen, G.S., Propp, S.S., Bromm, M., Rowland, K.J., Zhangm T., Rasper, D., Roy, S., Thornberry, N., Pinsky, L., Kaki-zuka, A., Ross, C.A., Nicholson, D.W., Bredesen, D.E. and Hayden, M.R. (1998). Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J. Biol. Chem.273, 9158–9167.

    Article  PubMed  CAS  Google Scholar 

  • Zegers, N.D., Claassen, E., Neelen, C, Mulder, E., van Laar, J.H., Voorhorst, M.M., Berrevoets, C.A., Brinkmann, A.O., van der Kwast, T.H., Ruizeveld de Winter, J.A. et al. (1991) Epitope prediction and confirmation for the human androgen receptor: generation of monoclonal antibodies for multi-assay performance following the synthetic peptide strategy. Biochim. Biophys. Acta1073, 23–32.

    PubMed  CAS  Google Scholar 

  • Zoghbi, H.Y. and Orr, H.T. (1999) Polyglutamine diseases: protein cleavage and Aggregation. Current Opinion in Neurobiology9, 566–570.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospitals, Toronto, Ontario, Canada

    Valerie Panet-Raymond, Bruce Gottlieb, Lenore K. Beitel, Hyman Schipper, Leonard Pinsky & Mark A. Trifiro

  2. Department of Biology, University of Toronto, Toronto, Ontario, Canada

    Valerie Panet-Raymond & Leonard Pinsky

  3. Department of Medicine, University of Toronto, Toronto, Ontario, Canada

    Hyman Schipper, Leonard Pinsky & Mark A. Trifiro

  4. Department of Pediatrics, University of Toronto, Toronto, Ontario, Canada

    Leonard Pinsky

  5. Department of Human Genetics, University of Toronto, Toronto, Ontario, Canada

    Leonard Pinsky & Mark A. Trifiro

  6. Department of Neurology and Neurosurgery, McGill University, University of Toronto, Toronto, Ontario, Canada

    Hyman Schipper

  7. Department of Biology, John Abbott Ste Anne de Bellevue, College, Quebec

    Bruce Gottlieb

  8. Division of Neurosurgery, University of Toronto, Toronto, Ontario, Canada

    Michael Timiansky

Authors
  1. Valerie Panet-Raymond
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  2. Bruce Gottlieb
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  3. Lenore K. Beitel
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  4. Hyman Schipper
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  5. Michael Timiansky
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  6. Leonard Pinsky
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  7. Mark A. Trifiro
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Corresponding author

Correspondence to Bruce Gottlieb.

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Panet-Raymond, V., Gottlieb, B., Beitel, L.K. et al. Characterization of intracellular aggregates using fluorescently-tagged polyglutamine-expanded androgen receptor. neurotox res 3, 259–275 (2001). https://doi.org/10.1007/BF03033265

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  • DOI: https://doi.org/10.1007/BF03033265

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