title: Naturally- and experimentally-designed restorations of the Parkin gene deficit in autosomal recessive juvenile parkinsonism.
abstract: Intranuclear events due to mutations in the Parkin gene remain elusive in autosomal recessive juvenile parkinsonism (ARJP). We identified a mutant PARKIN protein in fibroblast cultures from a pair of siblings with ARJP who were homozygous for the exon 4-deleted Parkin gene. Disease was mild in one patient and debilitating in the other. The detected mutant, encoded by a transcript lacking exon 3 as well as exon 4, is an in-frame deletion that removes 121 aa, resulting in a 344-aa protein (PaDel3,4). Cell culture and transfection studies revealed negative correlations between expression levels of PaDel3,4 and those of cell cycle proteins, including cyclin E, CDK2, ppRb, and E2F-1, and demonstrated that GFP-PaDel3,4 entered nucleus and ubiquitinated cyclin E as a part of SCF(hSel-10) ligase complex in the patient cells. In addition, nuclear localization signal-tagged PaDel3,4 expressed in the transfected patient cells most effectively ubiquitinated cyclin E and reduced DNA damage, protecting cells from oxidative stress. Antisense-oligonucleotide treatment promoted skipping of exon 3 and thus generated PaDel3,4, increasing cell survival. Collectively, we propose that naturally- and experimentally-induced exon skipping at least partly restores the mutant Parkin gene deficit, providing a molecular basis for the development of therapeutic exon skipping.
authors: Department of Neurology, Faculty of Medicine, Nara Medical University School of Medicine, Japan.
authors: Hirohide
authors: H
authors: Asai
authors: Makito
authors: M
authors: Hirano
authors: Takao
authors: T
authors: Kiriyama
authors: Masanori
authors: M
authors: Ikeda
authors: Satoshi
authors: S
authors: Ueno
mesh_terms: D000368:Aged; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002470:Cell Survival; D002478:Cells, Cultured; D019927:Cyclin E / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D005091:Exons / Q000235:genetics*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D017353:Gene Deletion; D006801:Humans; D008297:Male; D016376:Oligonucleotides, Antisense / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D018384:Oxidative Stress / Q000235:genetics; D020734:Parkinsonian Disorders / Q000235:genetics*; D044843:SKP Cullin F-Box Protein Ligases; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*
chemical_list: D019927:Cyclin E; D016376:Oligonucleotides, Antisense; D019328:Buthionine Sulfoximine; D044843:SKP Cullin F-Box Protein Ligases; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein