Title Page
Contents
초록 8
ABSTRACT 9
INTRODUCTION 10
MATERIALS AND METHODS 16
1. Cell culture and cell lines 16
2. Antibodies, chemicals, and Western blotting assay 16
3. In vivo ISGylation assay 17
4. Site-directed mutagenesis 18
5. Cycloheximide chase assay 18
RESULTS 20
1. NM23-H1 is a a target of ISG15 modification 20
2. USP18 removes ISG15 from NM23-H1 21
3. Lysine 49 and 66 of NM23-H1 are the major ISGylation site 21
4. NM23-H1 ISGylation increases NM23-H1 stability 22
5. NM23-H1 ISGylation increases p53 stability 23
6. Interaction of p53 and MDM2 is affected by NM23-H1 ISGylation 23
7. NM23-H1 ISGylation regulates Akt and Erk signaling 24
8. NM23-H1 ISGylation regulates EMT and CSC markers 25
9. NM23-H1 ISGylation regulates anti-tumorigenic activity in vitro 26
10. ISGylation status of NM23-H1 active site mutant 27
DISCUSSION 50
Reference 53
Fig 1. NM23-H1 is a target of ISG15 modification 28
Fig 2. USP18 removes ISG15 from NM23-H1 30
Fig 3. Lysine 49 and 66 of NM23-H1 are the major ISGylation site 32
Fig 4. NM23-H1 ISGylation increases NM23-H1 stability 34
Fig 5. NM23-H1 ISGylation increases p53 stability 36
Fig 6. Interaction of p53 and MDM2 is affected by NM23-H1 ISGylation 39
Fig 7. NM23-H1 ISGylation regulates Akt and Erk signaling 41
Fig 8. NM23-H1 ISGylation regulates EMT and CSC markers 43
Fig 9. NM23-H1 ISGylation regulates anti-tumorigenic activity in vitro 45
Fig 10. NM23-H1-active site mutant regulates ISGylation 48