MARCH5에 의한 RIG-I/MAVS 신호전달의 조절
Regulation of RIG-I/MAVS signaling by the MARCH5 ubiquitin ligase
- 주제(키워드) Innate immunity , Mitochondrial E3 ubiquitin ligase , MARCH5 , RIG-I , Oligomer , Aggregate
- 발행기관 아주대학교
- 지도교수 조혜성
- 발행년도 2019
- 학위수여년월 2019. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000028661
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The innate immune system has an important role in the mammalian immune response. Recent study has demonstrated that mitochondria take part in a broad range of innate immune pathways, functioning as signal platforms and resulting in activation of immune responses. Previously, our lab found that MARCH5 regulates immune response by resolving MAVS prion-like aggregates during viral infection. The present study revealed the kinetics of RIG-I/MAVS oligomerization upon poly(I:C) transfection. RIG-I and MAVS began to form oligomer, respectively after 12 hrs of poly(I:C) transfection. This was accompanied with accumulation of RIG-I and MAVS protein levels. At 36hrs after stimulation, the oligomerized RIG-I and MAVS proteins were accumulated in the NP_40-insoluble fractions, suggesting that the RIG-I/MAVS oligomers form a strong insoluble protein aggregates in cells. I found that MARCH5 not only targeted the oligomerized MAVS protein but also caused a significant reduction of the oligomerized RIG-I levels, of which degradation is mediated through a proteasome-dependent manner. In bone-marrow derived macrophage cell (BMDM) from MARCH5 knockout mice secreted high amount of type-I interferon secretion in response to poly(I:C) transfection. Approach to oligomeric complex provides understanding of relationship between RIG-I/MAVS oligomers and MARCH5. That make the reason I proposed that RIG-I and MAVS oligomers can be dual-targeted by MARCH5 during innate immune response.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 4
1. Plasmids and antibodies 4
2. Cell culture 4
3. Western blot analysis 5
4. Transfection of DNAs and siRNA 6
5. MAVS aggregate - insoluble fraction 6
6. Generation of MARCH5 conditional knock-out mouse. 7
III. RESULTS 9
1. RIG-I oligomer and MAVS aggregate increase upon poly(I:C) transfection 9
2. Increase of insoluble RIG-I oligomer after viral condition 12
3. The levels of IFN-beta and IL-6 increase in MARCH5 knock-out mouse upon virus challenge 14
4. MARCH5 negatively regulates oligomeric RIG-I 19
5. MARCH5 directly binds to RIG-I protein 23
6. RIG-I/MAVS complex formation is important for interaction with MARCH5 25
IV. DISCUSSION 27
V. REFERENCES 30
국문요약 36
