아주대학교

목차

I. INTRODUCTION 1
1. Autophagy 1
2. Formation of autophagosome 2
3. Autophagosome-lysosome fusion 3
4. Autophagy and neurodegenerative disease 5
5. TRIM proteins 7
6. The aims of this study 9
II. MATERIALS AND METHODS 10
1. DNA constructs 10
2. Antibodies 11
3. Chemicals and reagents 11
4. Immunoblotting 12
5. Immunoprecipitation 12
6. Cell culture 13
7. Plasmid transfection and RNA interference 13
8. Generation of stable cell lines 14
9. Immunofluorescence staining and confocal microscopy 14
10. Generation of knockout cell lines 15
11. Flow cytometry 16
12. Protein purification and in vitro pull-down assays 16
13. Quantification of colocalization and statistical analysis 17
14. Proximity ligation assay (PLA) 18
III. RESULTS 19
Chapter 1. TRIM22 facilitates autophagosome-lysosome fusion by mediating the association of GABARAPs and PLEKHM 1 19
1. TRIM22 deficiency interferes with intracellular clearance 19
2. TRIM22 positively regulates autophagosome-lysosome fusion 25
3. TRIM22 interacts with GABARAPs and mediates their lysosomal localization 35
4. TRIM22 positively regulates autophagosome-lysosome fusion by mediating the association of GABARAPs with PLEKHM1 45
5. TRIM22 facilitates autophagosome-lysosome fusion by interacting with PLEKHM 53
6. The function of TRIM22 as an E3 ubiquitin ligase is distinct from its role in the regulation of autophagy 59
7. AD-related TRIM22 variant interferes with intracellular clearance by inhibiting autophagosome-lysosome fusion 73
Chapter 2. TRIM22 promotes autophagosome formation by recruiting autophagy pre-initiation and initiation complexes 90
1. TRIM22 promotes autophagosome formation by recruiting ULK1 complex 90
2. TRIM22 promotes autophagosome formation by recruiting class III PI3K complex 102
IV. DISCUSSION 110
V. REFERENCES 119
국문요약 130