목차

I. INTRODUCTUON 1
II. MATERIALS AND METHOD 9
A. Plasmids and chemicals 9
B. Cell culture 9
C. Transfection and RNA interference 9
D. Generation of March5 KO RAW264.7 10
E. Western blot analysis 10
F. Mitochondrial fractionation 12
G. Pull-down assay 12
H. Ubiquitination assay 12
I. Immunofluorescence 13
J. Reverse transcription-quantitative polymerase chain reaction 13
K. Mitochondrial DNA copy number assay 15
L. ATP assay 15
M. Statistical analysis 16
III. RESULTS 17
A. Establishment of FokI endonuclease systems to generate mtDNA double-strand breaks　 17
B. MTERF1-FokI-mCherry and TFAM-FokI-mCherry localize to mitochondria　 19
C. Fluorescence signal of the mtDNA is reduced in an activity-dependent manner at the mt-FokI endonuclease　 21
D. Specific targeting of mtDNA by the FokI endonuclease reduces the amount of mtDNA　 23
E. Mt-FokIs do not affect nuclear DNA damage response and ATP synthesis upon mtDNA degradation　 26
F. A type I interferon response is triggered by mtDSBs even in the absence of cGAS　 29
G. Mitochondrial DNA cleavage increases phosphorylation of IRF3 and HSPA8 in a variety of cell lines　 32
H. DNA-PKcs mediates phosphorylation of HSPA8 and IRF3 in response to breaks in mtDNA　 34
I. Cleaved mtDNA elicits DNA PKcs-mediated type I interferon response via the VDAC macropore　 37
J. Mitochondrial E3 ligase MARCH5 degrades DNA-PKcs in a proteasome-dependent manner　 40
K. MARCH5 binds and ubiquitylates activated DNA-PKcs　 43
L. MARCH5 suppresses DNA-PKcs-mediated type I interferon response　 46
M. LPS treatment elicits DNA-PKcs-dependent type I interferon response in murine macrophages　 48
N. MARCH5 prevents prolonged type І Interferon response in murine macrophage stimulated by LPS　 51
IV. DISCUSSION　 54
REFERENCE　 57
국문요약　 64