title: The importance of gene dosage studies: mutational analysis of the parkin gene in early-onset parkinsonism.
abstract: Early-onset parkinsonism (EOP) may be associated with different mutations in the parkin gene, including exon deletions and duplications. To test for gene dosage alterations, we developed a new method of quantitative duplex PCR using the fluorescence resonance energy transfer technique on the LightCycler (Roche Diagnostics). In 21 patients with EOP, three mutations (a single base pair substitution in exon 3 and small deletions in exon 9) were detected by conventional mutational screening (single-strand conformation polymorphism and sequence analysis), while alterations of gene dosage were found in seven patients. We identified heterozygous and compound heterozygous deletions of exons 2, 3, 5 and 7. The latter was also found in the homozygous state. In addition, two heterozygous duplications of exon 4 were observed. Remarkably, two patients carried more than two parkin mutations. This is the first study systematically screening all 12 exons of parkin by real-time, kinetic quantification and clearly shows that mutational analysis of the parkin gene should include gene dosage studies. Furthermore, our method of quantitative PCR is easily applicable to any other gene to be screened for deletions or duplications of whole exons.
authors.affiliation: Department of Neurology, Medical University of Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Hedrich
authors.forename: M
authors.initials: M
authors.lastname: Kann
authors.forename: A J
authors.initials: AJ
authors.lastname: Lanthaler
authors.forename: A
authors.initials: A
authors.lastname: Dalski
authors.forename: C
authors.initials: C
authors.lastname: Eskelson
authors.forename: O
authors.initials: O
authors.lastname: Landt
authors.forename: E
authors.initials: E
authors.lastname: Schwinger
authors.forename: P
authors.initials: P
authors.lastname: Vieregge
authors.forename: A E
authors.initials: AE
authors.lastname: Lang
authors.forename: X O
authors.initials: XO
authors.lastname: Breakefield
authors.forename: L J
authors.initials: LJ
authors.lastname: Ozelius
authors.forename: P P
authors.initials: PP
authors.lastname: Pramstaller
authors.forename: C
authors.initials: C
authors.lastname: Klein
mesh_terms: D017668:Age of Onset; D015139:Blotting, Southern; D004252:DNA Mutational Analysis; D005260:Female; D018628:Gene Dosage; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008025:Ligases / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D020734:Parkinsonian Disorders / Q000235:genetics*; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods; D044767:Ubiquitin-Protein Ligases
chemical_list: D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D008025:Ligases