title: A mutant ataxin-3 putative-cleavage fragment in brains of Machado-Joseph disease patients and transgenic mice is cytotoxic above a critical concentration.
abstract: Machado-Joseph disease (MJD) is an inherited neurodegenerative disorder caused by ataxin-3 with a polyglutamine expansion. It is proposed that a toxic cleavage fragment of mutant ataxin-3 alternatively spliced isoform mjd1a triggers neurodegeneration, although this fragment has not yet been detected in the brains of MJD patients or in animal models. We have now generated transgenic mice expressing human mutant (Q71) or normal (Q20) ataxin-3 mjd1a under the control of the mouse prion promoter. Q71 transgenic mice expressing mutant ataxin-3 mjd1a above a critical level developed a phenotype similar to MJD including progressive postural instability, gait and limb ataxia, weight loss, premature death, neuronal intranuclear inclusions, and decreased tyrosine hydroxylase-positive neurons in the substantia nigra (determined by unbiased stereology). Q20 transgenic mice had normal behavior and pathology. Brains from sick Q71 transgenic mice contained an abundant mutant ataxin-3 mjd1a putative-cleavage fragment (Fragment), which was scarce in normal Q71 transgenic mice. Reactivity of the Fragment with a panel of antibodies and comigration with truncations of mutant ataxin-3 revealed that it contained residues C terminal to amino acid 221 to include the polyglutamine expansion. A similar portion of mutant ataxin-3 mjd1a expressed in transfected neuroblastoma cells was toxic above a critical concentration. The Fragment was more abundant in two affected brain regions of MJD patients. Thus, we have developed a murine model for mutant ataxin-3 mjd1a toxicity and identified a putative-cleavage fragment of the disease protein in the brains of these transgenic mice and MJD patients that is cytotoxic above a critical concentration.
authors.affiliation: Department of Psychiatry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Goti
authors.forename: Scott M
authors.initials: SM
authors.lastname: Katzen
authors.forename: Jesse
authors.initials: J
authors.lastname: Mez
authors.forename: Noam
authors.initials: N
authors.lastname: Kurtis
authors.forename: Jennifer
authors.initials: J
authors.lastname: Kiluk
authors.forename: Lea
authors.initials: L
authors.lastname: Ben-Haïem
authors.forename: Nancy A
authors.initials: NA
authors.lastname: Jenkins
authors.forename: Neal G
authors.initials: NG
authors.lastname: Copeland
authors.forename: Akira
authors.initials: A
authors.lastname: Kakizuka
authors.forename: Alan H
authors.initials: AH
authors.lastname: Sharp
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Ross
authors.forename: Peter R
authors.initials: PR
authors.lastname: Mouton
authors.forename: Veronica
authors.initials: V
authors.lastname: Colomer
mesh_terms: D000328:Adult; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000067699:Ataxin-3; D001923:Brain Chemistry; D045744:Cell Line, Tumor / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005106:Exploratory Behavior; D005260:Female; D018737:Hand Strength; D006801:Humans; D045586:Intranuclear Inclusion Bodies; D017827:Machado-Joseph Disease / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008875:Middle Aged; D018598:Minisatellite Repeats; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D009447:Neuroblastoma / Q000473:pathology; D009474:Neurons / Q000473:pathology; D009687:Nuclear Proteins; D010446:Peptide Fragments / Q000032:analysis / Q000633:toxicity*; D010641:Phenotype; D004856:Postural Balance; D012018:Reflex; D012097:Repressor Proteins; D013347:Subcellular Fractions; D013378:Substantia Nigra / Q000473:pathology; D014157:Transcription Factors; D014162:Transfection
chemical_list: D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D012097:Repressor Proteins; D014157:Transcription Factors; C092341:ATXN3 protein, human; D000067699:Ataxin-3; C511594:Atxn3 protein, mouse