title: Mutant torsinA, which causes early-onset primary torsion dystonia, is redistributed to membranous structures enriched in vesicular monoamine transporter in cultured human SH-SY5Y cells.
abstract: A single GAG deletion in the DYT1 gene causes primary early-onset, generalized torsion dystonia. The DYT1 protein product, torsinA, belongs to the AAA+ family of proteins. When overexpressed, wild-type torsinA localizes mainly to the endoplasmic reticulum, whereas the mutant forms inclusions of unclear biogenetic origin. In this study, overexpressed wild-type torsinA in human neuroblastoma (SH-SY5Y) cell lines was distributed throughout the cell body and colocalized with a marker for the endoplasmic reticulum, confirming it is an endoplasmic reticulum protein. However, mutant torsinA showed perinuclear staining and formed distinct globular inclusions, which did not colocalize with endoplasmic reticulum markers. Immunoelectron microscopy of the mutant torsinA inclusions revealed membrane whorls staining for torsinA, as well as labeling of lamellae, isolated bilayers, and perinuclear membranes. This finding shows that mutant torsinA redistributes to specific membranous structures, which may represent different stages of maturation of the intracellular inclusions. The mutant torsinA-containing bodies were immunoreactive for vesicular monoamine transporter 2 (VMAT2). VMAT2 expression is important for the exocytosis of bioactive monoamines in neurons. Abnormal processing, transport, or entrapment of VMAT2 within the mutant torsinA membranous inclusions, therefore, may affect cellular dopamine release, providing a potential pathogenic mechanism for the DYT1-dependent dystonia.
authors.affiliation: Department of Clinical Neurosciences, Royal Free and University College Medical School, London, United Kingdom.
authors.forename: Anjum
authors.initials: A
authors.lastname: Misbahuddin
authors.affiliation: Department of Clinical Neurosciences, Royal Free and University College Medical School, London, United Kingdom.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Placzek
authors.affiliation: Department of Clinical Neurosciences, Royal Free and University College Medical School, London, United Kingdom.
authors.forename: Jan-Willem
authors.initials: JW
authors.lastname: Taanman
authors.affiliation: Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, London, United Kingdom.
authors.forename: Steve
authors.initials: S
authors.lastname: Gschmeissner
authors.affiliation: Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, London, United Kingdom.
authors.forename: Giampietro
authors.initials: G
authors.lastname: Schiavo
authors.affiliation: Department of Clinical Neurosciences, Royal Free and University College Medical School, London, United Kingdom.
authors.forename: J Mark
authors.initials: JM
authors.lastname: Cooper
authors.affiliation: Department of Clinical Neurosciences, Royal Free and University College Medical School, London, United Kingdom.
authors.forename: Thomas T
authors.initials: TT
authors.lastname: Warner
mesh_terms: D000367:Age Factors; D015153:Blotting, Western; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000235:genetics; D004422:Dystonia Musculorum Deformans / Q000235:genetics* / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D008854:Microscopy, Electron; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism*; D009447:Neuroblastoma / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D017354:Point Mutation / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D018911:Trinucleotide Repeats / Q000235:genetics; D050492:Vesicular Biogenic Amine Transport Proteins; D050493:Vesicular Monoamine Transport Proteins
chemical_list: D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D018832:Molecular Chaperones; C493452:SLC18A2 protein, human; C108175:TOR1A protein, human; D050492:Vesicular Biogenic Amine Transport Proteins; D050493:Vesicular Monoamine Transport Proteins