Genetic analysis of PARK2 gene in Korean patients with Parkinson's disease

다국어 초록 (Multilingual Abstract)

Mutations in PARK2 are considered a common cause of Parkinson’s disease (PD). To assess the frequency of PARK2 mutations in the Korean population, we screened the PARK2 gene in 83 Korean PD patients: two young onset (YO, ≤49), 32 middle onset (MO, 50-69) and 49 late onset (LO, 70). Detection of the point mutations was performed by direct sequencing of the PARK2 exons, and exonic rearrangements were analyzed using multiplex ligation-dependent probe amplification (MLPA). Five known PARK2 variants were identified in 53 (63.9%) of the Korean PD patients: two missense mutations (Y267H and M458L) and three polymorphisms (S167N, L272I and V380L). We also found an increased frequency of PARK2 variants in PD patients and a lowered PD age at onset (AAO) in those having two variants, suggesting that the genetic variation in PARK2 gene might be a genetic risk factor of PD in Korean population.

목차 (Table of Contents)

• ▣ PART I
• I-A. ABSTRACT 1
• I-B. INTRODUCTION 2
• I-C. MATERIALS AND METHODS 4
• I-C-1. Subjects 4
• I-C-2. Mutation analysis 4
• I-C-3. Gene dosage analysis 5
• I-C-4. Statistical evaluation 5
• I-D. RESULTS 6
• I-D-1. Frequency of Parkin mutations 6
• I-D-2. Exon dosages analysis of Parkin 7
• I-E. DISCUSSION 8
• I-F. REFERENCES 11
• ▣ PART II
• II-A. ABSTRACT 29
• II-B. INTRODUCTION 30
• II-C. MATERIALS AND METHODS 32
• II-C-1. Cell culture and transfection 32
• II-C-2. Plasmids construction and mutagenesis 32
• II-C-3. Antibodies 33
• II-C-4. Glutathione S-transferase (GST) protein preparation 33
• II-C-5. In vitro auto-ubiquitination assay 34
• II-C-6. In cell auto-ubiquitination assay 34
• II-D. RESULTS 35
• II-D-1. PARK2 M458L was identified in Korean PD patient 35
• II-D-2. PARK2 M458L impairs PARK2-mediated degradation of
• substrates (AIMP2 and SNCA) 36
• II-D-3. PARK2 M458L mediates E2-independent ubiquitination
• in vitro 37
• II-D-4. PARK2 M458L impairs its enzymatic function in living cells 38
• II-E. DISCUSSION 39
• II-F. REFERENCES 41
• III. SUMMARY (IN KOREAN) 53