Self-assembly of polyglutamine-containing huntingtin fragments into amyloid-like fibrils: implications for Huntington's disease pathology.

Huntington's disease is a progressive neurodegenerative disorder caused by a polyglutamine [poly(Q)] repeat expansion in the first exon of the huntingtin protein. Previously, we showed that N-terminal huntingtin peptides with poly(Q) tracts in the pathological range (51-122 glutamines), but not with poly(Q) tracts in the normal range (20 and 30 glutamines), form high molecular weight protein aggregates with a fibrillar or ribbon-like morphology, reminiscent of scrapie prion rods and beta-amyloid fibrils in Alzheimer's disease. Here we report that the formation of amyloid-like huntingtin aggregates in vitro not only depends on poly(Q) repeat length but also critically depends on protein concentration and time. Furthermore, the in vitro aggregation of huntingtin can be seeded by preformed fibrils. Together, these results suggest that amyloid fibrillogenesis in Huntington's disease, like in Alzheimer's disease, is a nucleation-dependent polymerization.

[1]  P. Fraser,et al.  Amyloid-like properties of a synthetic peptide corresponding to the carboxy terminus of beta-amyloid protein precursor. , 1992, Archives of biochemistry and biophysics.

[2]  Harry T Orr,et al.  Ataxin-1 Nuclear Localization and Aggregation Role in Polyglutamine-Induced Disease in SCA1 Transgenic Mice , 1998, Cell.

[3]  J. Mandel,et al.  A cellular model that recapitulates major pathogenic steps of Huntington's disease. , 1998, Human molecular genetics.

[4]  M. Perutz,et al.  Glutamine repeats and inherited neurodegenerative diseases: molecular aspects. , 1996, Current opinion in structural biology.

[5]  H. Paulson,et al.  Recruitment and the Role of Nuclear Localization in Polyglutamine-mediated Aggregation , 1998, The Journal of cell biology.

[6]  K. Fischbeck,et al.  Intranuclear Inclusions of Expanded Polyglutamine Protein in Spinocerebellar Ataxia Type 3 , 1997, Neuron.

[7]  D. Smith,et al.  Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. , 1988, Gene.

[8]  L. J. Fisher.  Neural Precursor Cells: Applications for the Study and Repair of the Central Nervous System , 1997, Neurobiology of Disease.

[9]  S. N. Timasheff,et al.  The measurement of cooperative protein self-assembly by turbidity and other techniques. , 1986, Methods in enzymology.

[10]  H. Lehrach,et al.  SH3GL3 associates with the Huntingtin exon 1 protein and promotes the formation of polygln-containing protein aggregates. , 1998, Molecular cell.

[11]  J. Hofrichter,et al.  The biophysics of sickle cell hydroxyurea therapy. , 1995, Science.

[12]  Manish S. Shah,et al.  A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes , 1993, Cell.

[13]  S. W. Davies,et al.  Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. , 1997, Science.

[14]  S. Tsuji,et al.  Suppression of aggregate formation and apoptosis by transglutaminase inhibitors in cells expressing truncated DRPLA protein with an expanded polyglutamine stretch , 1998, Nature Genetics.

[15]  H. Zoghbi,et al.  Ataxin-1 with an expanded glutamine tract alters nuclear matrix-associated structures , 1997, Nature.

[16]  P. Chambon,et al.  Structure, localization and transcriptional properties of two classes of retinoic acid receptor alpha fusion proteins in acute promyelocytic leukemia (APL): structural similarities with a new family of oncoproteins. , 1992, The EMBO journal.

[17]  K. Kosik,et al.  The Alzheimer's disease sphinx: a riddle with plaques and tangles , 1994, The Journal of cell biology.

[18]  P. Lansbury,et al.  Seeding “one-dimensional crystallization” of amyloid: A pathogenic mechanism in Alzheimer's disease and scrapie? , 1993, Cell.

[19]  G. Schellenberg,et al.  Genetic dissection of Alzheimer disease, a heterogeneous disorder. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Hans Lehrach,et al.  Huntingtin-Encoded Polyglutamine Expansions Form Amyloid-like Protein Aggregates In Vitro and In Vivo , 1997, Cell.

[21]  P. Lansbury,et al.  Models of amyloid seeding in Alzheimer's disease and scrapie: mechanistic truths and physiological consequences of the time-dependent solubility of amyloid proteins. , 1997, Annual review of biochemistry.

[22]  Claudio Soto,et al.  β-sheet breaker peptides inhibit fibrillogenesis in a rat brain model of amyloidosis: Implications for Alzheimer's therapy , 1998, Nature Medicine.

[23]  K. Sathasivam,et al.  Identification of an HD patient with a (CAG)180 repeat expansion and the propagation of highly expanded CAG repeats in lambda phage , 1997, Human Genetics.

[24]  Steven Finkbeiner,et al.  Huntingtin Acts in the Nucleus to Induce Apoptosis but Death Does Not Correlate with the Formation of Intranuclear Inclusions , 1998, Cell.

[25]  Carl W. Cotman,et al.  Neurodegeneration induced by beta-amyloid peptides in vitro: the role of peptide assembly state , 1993, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[26]  B. Chesebro,et al.  Prion protein and the transmissible spongiform encephalopathies. , 1997, Trends in cell biology.

[27]  A Dürr,et al.  Spinocerebellar ataxia type 7 (SCA7): a neurodegenerative disorder with neuronal intranuclear inclusions. , 1998, Human molecular genetics.

[28]  Fumiaki Tanaka,et al.  Nuclear inclusions of the androgen receptor protein in spinal and bulbar muscular atrophy , 1998, Annals of neurology.

[29]  P S Harper,et al.  Phenotypic characterization of individuals with 30-40 CAG repeats in the Huntington disease (HD) gene reveals HD cases with 36 repeats and apparently normal elderly individuals with 36-39 repeats. , 1996, American journal of human genetics.

[30]  Mark Turmaine,et al.  Formation of Neuronal Intranuclear Inclusions Underlies the Neurological Dysfunction in Mice Transgenic for the HD Mutation , 1997, Cell.