Intracellular green fluorescent protein-polyalanine aggregates are associated with cell death.

Eight diseases, exemplified by Huntington's disease and spinocerebellar ataxia type 1, are caused by CAG-repeat expansion mutations. The CAG repeats are translated into expanded polyglutamine tracts, which are associated with deleterious novel functions. While these diseases are characterized by intraneuronal aggregate formation, it is unclear whether the aggregates cause disease. We have addressed this debate by generating intracellular aggregates with green fluorescent protein (GFP) fused to 19-37 alanines. No aggregates were seen in cells expressing native GFP or GFP fused to seven alanines. Aggregate-containing cells expressing GFP fused to 19-37 polyalanines show high rates of nuclear fragmentation compared with cells expressing the same constructs without aggregates, or cells expressing GFP fused to seven alanines. This suggests an association between aggregate formation and cell death.

[1]  O. Quarrell.  Glutamine repeats and neurodegenerative diseases: molecular aspects , 2001, Human Genetics.

[2]  H. Paulson,et al.  Analysis of the Role of Heat Shock Protein (Hsp) Molecular Chaperones in Polyglutamine Disease , 1999, The Journal of Neuroscience.

[3]  Harry T Orr,et al.  Mutation of the E6-AP Ubiquitin Ligase Reduces Nuclear Inclusion Frequency While Accelerating Polyglutamine-Induced Pathology in SCA1 Mice , 1999, Neuron.

[4]  T. Uchihara,et al.  Neuronal intranuclear inclusions in spinocerebellar ataxia type 2: triple-labeling immunofluorescent study , 1999, Neuroscience Letters.

[5]  D. Housman,et al.  Insoluble detergent-resistant aggregates form between pathological and nonpathological lengths of polyglutamine in mammalian cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[6]  I. Kanazawa,et al.  Abundant expression and cytoplasmic aggregations of [alpha]1A voltage-dependent calcium channel protein associated with neurodegeneration in spinocerebellar ataxia type 6. , 1999, Human molecular genetics.

[7]  S. Tsuji,et al.  Adenovirus-mediated expression of mutant DRPLA proteins with expanded polyglutamine stretches in neuronally differentiated PC12 cells. Preferential intranuclear aggregate formation and apoptosis. , 1999, Human molecular genetics.

[8]  D. Rubinsztein,et al.  Intracellular inclusions, pathological markers in diseases caused by expanded polyglutamine tracts? , 1999, Journal of medical genetics.

[9]  M. MacDonald,et al.  Amyloid Formation by Mutant Huntingtin: Threshold, Progressivity and Recruitment of Normal Polyglutamine Proteins , 1998, Somatic cell and molecular genetics.

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

[11]  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.

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

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

[14]  C A Ross,et al.  Truncated N-terminal fragments of huntingtin with expanded glutamine repeats form nuclear and cytoplasmic aggregates in cell culture. , 1998, Human molecular genetics.

[15]  J. Rommens,et al.  Short GCG expansions in the PABP2 gene cause oculopharyngeal muscular dystrophy , 1998, Nature Genetics.

[16]  A. Hackam,et al.  Length of huntingtin and its polyglutamine tract influences localization and frequency of intracellular aggregates , 1998, Nature Genetics.

[17]  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.

[18]  S. W. Davies,et al.  Intranuclear Neuronal Inclusions in Huntington's Disease and Dentatorubral and Pallidoluysian Atrophy: Correlation between the Density of Inclusions andIT15CAG Triplet Repeat Length , 1998, Neurobiology of Disease.

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

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

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

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

[23]  R. Houghten,et al.  Polyalanine-based peptides as models for self-associated beta-pleated-sheet complexes. , 1997, Biochemistry.

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