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Philosophical Transactions of the Royal Society B: Biological Sciences logoLink to Philosophical Transactions of the Royal Society B: Biological Sciences
. 1999 Jun 29;354(1386):1047–1055. doi: 10.1098/rstb.1999.0457

Evidence for both the nucleus and cytoplasm as subcellular sites of pathogenesis in Huntington's disease in cell culture and in transgenic mice expressing mutant huntingtin.

A S Hackam 1, J G Hodgson 1, R Singaraja 1, T Zhang 1, L Gan 1, C A Gutekunst 1, S M Hersch 1, M R Hayden 1
PMCID: PMC1692613  PMID: 10434304

Abstract

A unifying feature of the CAG expansion diseases is the formation of intracellular aggregates composed of the mutant polyglutamine-expanded protein. Despite the presence of aggregates in affected patients, the precise relationship between aggregates and disease pathogenesis is unresolved. Results from in vivo and in vitro studies of mutant huntingtin have led to the hypothesis that nuclear localization of aggregates is critical for the pathology of Huntington's disease (HD). We tested this hypothesis using a 293T cell culture model system by comparing the frequency and toxicity of cytoplasmic and nuclear huntingtin aggregates. Insertion of nuclear import or export sequences into huntingtin fragments containing 548 or 151 amino acids was used to reverse the normal localization of these proteins. Changing the subcellular localization of the fragments did not influence their total aggregate frequency. There were also no significant differences in toxicity associated with the presence of nuclear compared with cytoplasmic aggregates. These studies, together with findings in transgenic mice, suggest two phases for the pathogenesis of HD, with the initial toxicity in the cytoplasm followed by proteolytic processing of huntingtin, nuclear translocation with increased nuclear concentration of N-terminal fragments, seeding of aggregates and resultant apoptotic death. These findings support the nucleus and cytosol as subcellular sites for pathogenesis in HD.

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Selected References

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  1. Andrew S. E., Goldberg Y. P., Hayden M. R. Rethinking genotype and phenotype correlations in polyglutamine expansion disorders. Hum Mol Genet. 1997 Nov;6(12):2005–2010. doi: 10.1093/hmg/6.12.2005. [DOI] [PubMed] [Google Scholar]
  2. Becher M. W., Kotzuk J. A., Sharp A. H., Davies S. W., Bates G. P., Price D. L., Ross C. A. Intranuclear neuronal inclusions in Huntington's disease and dentatorubral and pallidoluysian atrophy: correlation between the density of inclusions and IT15 CAG triplet repeat length. Neurobiol Dis. 1998 Apr;4(6):387–397. doi: 10.1006/nbdi.1998.0168. [DOI] [PubMed] [Google Scholar]
  3. Bots G. T., Bruyn G. W. Neuropathological changes of the nucleus accumbens in Huntington's chorea. Acta Neuropathol. 1981;55(1):21–22. doi: 10.1007/BF00691525. [DOI] [PubMed] [Google Scholar]
  4. Burke J. R., Enghild J. J., Martin M. E., Jou Y. S., Myers R. M., Roses A. D., Vance J. M., Strittmatter W. J. Huntingtin and DRPLA proteins selectively interact with the enzyme GAPDH. Nat Med. 1996 Mar;2(3):347–350. doi: 10.1038/nm0396-347. [DOI] [PubMed] [Google Scholar]
  5. Butler R., Leigh P. N., McPhaul M. J., Gallo J. M. Truncated forms of the androgen receptor are associated with polyglutamine expansion in X-linked spinal and bulbar muscular atrophy. Hum Mol Genet. 1998 Jan;7(1):121–127. doi: 10.1093/hmg/7.1.121. [DOI] [PubMed] [Google Scholar]
  6. Carmichael J., DeGraff W. G., Gazdar A. F., Minna J. D., Mitchell J. B. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 1987 Feb 15;47(4):936–942. [PubMed] [Google Scholar]
  7. 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. Truncated N-terminal fragments of huntingtin with expanded glutamine repeats form nuclear and cytoplasmic aggregates in cell culture. Hum Mol Genet. 1998 May;7(5):783–790. doi: 10.1093/hmg/7.5.783. [DOI] [PubMed] [Google Scholar]
  8. Cummings C. J., Mancini M. A., Antalffy B., DeFranco D. B., Orr H. T., Zoghbi H. Y. Chaperone suppression of aggregation and altered subcellular proteasome localization imply protein misfolding in SCA1. Nat Genet. 1998 Jun;19(2):148–154. doi: 10.1038/502. [DOI] [PubMed] [Google Scholar]
  9. Davies S. W., Turmaine M., Cozens B. A., DiFiglia M., Sharp A. H., Ross C. A., Scherzinger E., Wanker E. E., Mangiarini L., Bates G. P. Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell. 1997 Aug 8;90(3):537–548. doi: 10.1016/s0092-8674(00)80513-9. [DOI] [PubMed] [Google Scholar]
  10. DiFiglia M., Sapp E., Chase K. O., Davies S. W., Bates G. P., Vonsattel J. P., Aronin N. Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science. 1997 Sep 26;277(5334):1990–1993. doi: 10.1126/science.277.5334.1990. [DOI] [PubMed] [Google Scholar]
  11. Ellerby L. M., Andrusiak R. L., Wellington C. L., Hackam A. S., Propp S. S., Wood J. D., Sharp A. H., Margolis R. L., Ross C. A., Salvesen G. S. Cleavage of atrophin-1 at caspase site aspartic acid 109 modulates cytotoxicity. J Biol Chem. 1999 Mar 26;274(13):8730–8736. doi: 10.1074/jbc.274.13.8730. [DOI] [PubMed] [Google Scholar]
  12. Ellerby L. M., Hackam A. S., Propp S. S., Ellerby H. M., Rabizadeh S., Cashman N. R., Trifiro M. A., Pinsky L., Wellington C. L., Salvesen G. S. Kennedy's disease: caspase cleavage of the androgen receptor is a crucial event in cytotoxicity. J Neurochem. 1999 Jan;72(1):185–195. doi: 10.1046/j.1471-4159.1999.0720185.x. [DOI] [PubMed] [Google Scholar]
  13. Goldberg Y. P., 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. Cleavage of huntingtin by apopain, a proapoptotic cysteine protease, is modulated by the polyglutamine tract. Nat Genet. 1996 Aug;13(4):442–449. doi: 10.1038/ng0896-442. [DOI] [PubMed] [Google Scholar]
  14. Gourfinkel-An I., Cancel G., Duyckaerts C., Faucheux B., Hauw J. J., Trottier Y., Brice A., Agid Y., Hirsch E. C. Neuronal distribution of intranuclear inclusions in Huntington's disease with adult onset. Neuroreport. 1998 Jun 1;9(8):1823–1826. doi: 10.1097/00001756-199806010-00028. [DOI] [PubMed] [Google Scholar]
  15. Green D. R., Reed J. C. Mitochondria and apoptosis. Science. 1998 Aug 28;281(5381):1309–1312. doi: 10.1126/science.281.5381.1309. [DOI] [PubMed] [Google Scholar]
  16. Görlich D., Mattaj I. W. Nucleocytoplasmic transport. Science. 1996 Mar 15;271(5255):1513–1518. doi: 10.1126/science.271.5255.1513. [DOI] [PubMed] [Google Scholar]
  17. Hackam A. S., Singaraja R., Wellington C. L., Metzler M., McCutcheon K., Zhang T., Kalchman M., Hayden M. R. The influence of huntingtin protein size on nuclear localization and cellular toxicity. J Cell Biol. 1998 Jun 1;141(5):1097–1105. doi: 10.1083/jcb.141.5.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hackam A. S., Singaraja R., Zhang T., Gan L., Hayden M. R. In vitro evidence for both the nucleus and cytoplasm as subcellular sites of pathogenesis in Huntington's disease. Hum Mol Genet. 1999 Jan;8(1):25–33. doi: 10.1093/hmg/8.1.25. [DOI] [PubMed] [Google Scholar]
  19. Hackam A. S., Wellington C. L., Hayden M. R. The fatal attraction of polyglutamine-containing proteins. Clin Genet. 1998 Apr;53(4):233–242. doi: 10.1111/j.1399-0004.1998.tb02687.x. [DOI] [PubMed] [Google Scholar]
  20. Holmberg M., Duyckaerts C., Dürr A., Cancel G., Gourfinkel-An I., Damier P., Faucheux B., Trottier Y., Hirsch E. C., Agid Y. Spinocerebellar ataxia type 7 (SCA7): a neurodegenerative disorder with neuronal intranuclear inclusions. Hum Mol Genet. 1998 May;7(5):913–918. doi: 10.1093/hmg/7.5.913. [DOI] [PubMed] [Google Scholar]
  21. Igarashi S., Koide R., Shimohata T., Yamada M., Hayashi Y., Takano H., Date H., Oyake M., Sato T., Sato A. Suppression of aggregate formation and apoptosis by transglutaminase inhibitors in cells expressing truncated DRPLA protein with an expanded polyglutamine stretch. Nat Genet. 1998 Feb;18(2):111–117. doi: 10.1038/ng0298-111. [DOI] [PubMed] [Google Scholar]
  22. Kalchman M. A., Koide H. B., McCutcheon K., Graham R. K., Nichol K., Nishiyama K., Kazemi-Esfarjani P., Lynn F. C., Wellington C., Metzler M. HIP1, a human homologue of S. cerevisiae Sla2p, interacts with membrane-associated huntingtin in the brain. Nat Genet. 1997 May;16(1):44–53. doi: 10.1038/ng0597-44. [DOI] [PubMed] [Google Scholar]
  23. Klement I. A., Skinner P. J., Kaytor M. D., Yi H., Hersch S. M., Clark H. B., Zoghbi H. Y., Orr H. T. Ataxin-1 nuclear localization and aggregation: role in polyglutamine-induced disease in SCA1 transgenic mice. Cell. 1998 Oct 2;95(1):41–53. doi: 10.1016/s0092-8674(00)81781-x. [DOI] [PubMed] [Google Scholar]
  24. Li M., Nakagomi Y., Kobayashi Y., Merry D. E., Tanaka F., Doyu M., Mitsuma T., Hashizume Y., Fischbeck K. H., Sobue G. Nonneural nuclear inclusions of androgen receptor protein in spinal and bulbar muscular atrophy. Am J Pathol. 1998 Sep;153(3):695–701. doi: 10.1016/S0002-9440(10)65612-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Li S. H., Li X. J. Aggregation of N-terminal huntingtin is dependent on the length of its glutamine repeats. Hum Mol Genet. 1998 May;7(5):777–782. doi: 10.1093/hmg/7.5.777. [DOI] [PubMed] [Google Scholar]
  26. Li X. J., Li S. H., Sharp A. H., Nucifora F. C., Jr, Schilling G., Lanahan A., Worley P., Snyder S. H., Ross C. A. A huntingtin-associated protein enriched in brain with implications for pathology. Nature. 1995 Nov 23;378(6555):398–402. doi: 10.1038/378398a0. [DOI] [PubMed] [Google Scholar]
  27. Lunkes A., Mandel J. L. A cellular model that recapitulates major pathogenic steps of Huntington's disease. Hum Mol Genet. 1998 Sep;7(9):1355–1361. doi: 10.1093/hmg/7.9.1355. [DOI] [PubMed] [Google Scholar]
  28. Lunkes A., Mandel J. L. Polyglutamines, nuclear inclusions and neurodegeneration. Nat Med. 1997 Nov;3(11):1201–1202. doi: 10.1038/nm1197-1201. [DOI] [PubMed] [Google Scholar]
  29. Martindale D., Hackam A., Wieczorek A., Ellerby L., Wellington C., McCutcheon K., Singaraja R., Kazemi-Esfarjani P., Devon R., Kim S. U. Length of huntingtin and its polyglutamine tract influences localization and frequency of intracellular aggregates. Nat Genet. 1998 Feb;18(2):150–154. doi: 10.1038/ng0298-150. [DOI] [PubMed] [Google Scholar]
  30. Matilla A., Koshy B. T., Cummings C. J., Isobe T., Orr H. T., Zoghbi H. Y. The cerebellar leucine-rich acidic nuclear protein interacts with ataxin-1. Nature. 1997 Oct 30;389(6654):974–978. doi: 10.1038/40159. [DOI] [PubMed] [Google Scholar]
  31. Merry D. E., Kobayashi Y., Bailey C. K., Taye A. A., Fischbeck K. H. Cleavage, aggregation and toxicity of the expanded androgen receptor in spinal and bulbar muscular atrophy. Hum Mol Genet. 1998 Apr;7(4):693–701. doi: 10.1093/hmg/7.4.693. [DOI] [PubMed] [Google Scholar]
  32. 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. Ectopically expressed CAG repeats cause intranuclear inclusions and a progressive late onset neurological phenotype in the mouse. Cell. 1997 Dec 12;91(6):753–763. doi: 10.1016/s0092-8674(00)80464-x. [DOI] [PubMed] [Google Scholar]
  33. Paulson H. L., Perez M. K., Trottier Y., Trojanowski J. Q., Subramony S. H., Das S. S., Vig P., Mandel J. L., Fischbeck K. H., Pittman R. N. Intranuclear inclusions of expanded polyglutamine protein in spinocerebellar ataxia type 3. Neuron. 1997 Aug;19(2):333–344. doi: 10.1016/s0896-6273(00)80943-5. [DOI] [PubMed] [Google Scholar]
  34. Ross C. A. Intranuclear neuronal inclusions: a common pathogenic mechanism for glutamine-repeat neurodegenerative diseases? Neuron. 1997 Dec;19(6):1147–1150. doi: 10.1016/s0896-6273(00)80405-5. [DOI] [PubMed] [Google Scholar]
  35. Sapp E., Schwarz C., Chase K., Bhide P. G., Young A. B., Penney J., Vonsattel J. P., Aronin N., DiFiglia M. Huntingtin localization in brains of normal and Huntington's disease patients. Ann Neurol. 1997 Oct;42(4):604–612. doi: 10.1002/ana.410420411. [DOI] [PubMed] [Google Scholar]
  36. Saudou F., Finkbeiner S., Devys D., Greenberg M. E. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. Cell. 1998 Oct 2;95(1):55–66. doi: 10.1016/s0092-8674(00)81782-1. [DOI] [PubMed] [Google Scholar]
  37. Skinner P. J., Koshy B. T., Cummings C. J., Klement I. A., Helin K., Servadio A., Zoghbi H. Y., Orr H. T. Ataxin-1 with an expanded glutamine tract alters nuclear matrix-associated structures. Nature. 1997 Oct 30;389(6654):971–974. doi: 10.1038/40153. [DOI] [PubMed] [Google Scholar]
  38. Stevanin G., Trottier Y., Cancel G., Dürr A., David G., Didierjean O., Bürk K., Imbert G., Saudou F., Abada-Bendib M. Screening for proteins with polyglutamine expansions in autosomal dominant cerebellar ataxias. Hum Mol Genet. 1996 Dec;5(12):1887–1892. doi: 10.1093/hmg/5.12.1887. [DOI] [PubMed] [Google Scholar]
  39. Tait D., Riccio M., Sittler A., Scherzinger E., Santi S., Ognibene A., Maraldi N. M., Lehrach H., Wanker E. E. Ataxin-3 is transported into the nucleus and associates with the nuclear matrix. Hum Mol Genet. 1998 Jun;7(6):991–997. doi: 10.1093/hmg/7.6.991. [DOI] [PubMed] [Google Scholar]
  40. Tellez-Nagel I., Johnson A. B., Terry R. D. Studies on brain biopsies of patients with Huntington's chorea. J Neuropathol Exp Neurol. 1974 Apr;33(2):308–332. doi: 10.1097/00005072-197404000-00008. [DOI] [PubMed] [Google Scholar]
  41. Trottier Y., Lutz Y., Stevanin G., Imbert G., Devys D., Cancel G., Saudou F., Weber C., David G., Tora L. Polyglutamine expansion as a pathological epitope in Huntington's disease and four dominant cerebellar ataxias. Nature. 1995 Nov 23;378(6555):403–406. doi: 10.1038/378403a0. [DOI] [PubMed] [Google Scholar]
  42. Warrick J. M., Paulson H. L., Gray-Board G. L., Bui Q. T., Fischbeck K. H., Pittman R. N., Bonini N. M. Expanded polyglutamine protein forms nuclear inclusions and causes neural degeneration in Drosophila. Cell. 1998 Jun 12;93(6):939–949. doi: 10.1016/s0092-8674(00)81200-3. [DOI] [PubMed] [Google Scholar]
  43. 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. Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J Biol Chem. 1998 Apr 10;273(15):9158–9167. doi: 10.1074/jbc.273.15.9158. [DOI] [PubMed] [Google Scholar]

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