| 摘要 | 第6-7页 |
| abstract | 第7页 |
| Abbreviations | 第11-14页 |
| CHAPTER 1 INTRODUCTION | 第14-31页 |
| 1.1 Restriction factors | 第14-22页 |
| 1.1.1 Discovery and molecular biology of SERINC genes | 第17-18页 |
| 1.1.2 Identification of Ser5 and Ser3 as restriction factors for HIV- | 第18-19页 |
| 1.1.3 Ser5 mediated inhibition of viral infectivity | 第19-21页 |
| 1.1.4 Mechanistic understanding of viral fusion inhibition triggered by Ser | 第21-22页 |
| 1.2 Retroviruses | 第22-30页 |
| 1.2.1 HIV-1 | 第24-27页 |
| 1.2.2 MLV glycoGag | 第27-28页 |
| 1.2.3 EIAV S2 | 第28-30页 |
| 1.3 Aims of the project | 第30-31页 |
| CHAPTER 2 MURINE LEUKEMIA VIRUS ACCESSORY PROTEINS GLYCOGAG REDUCES MURINE SERINC5 PROTEIN EXPRESSION LEVEL VIA DEGRADING LYSOSOMAL PATHWAY TO COUNTERACT SERINC5 ANTIRETROVIRAL ACTIVITY | 第31-59页 |
| 2.1 Background | 第31-32页 |
| 2.2 Material and methods | 第32-38页 |
| 2.2.1 Cells and culturing system | 第32页 |
| 2.2.2 Plasmids | 第32-34页 |
| 2.2.3 Transfection | 第34-35页 |
| 2.2.4 Effect of mSer5 on HIV-1 infectivity | 第35页 |
| 2.2.5 Effect of mSer5 on MLV infectivity | 第35页 |
| 2.2.6 Effect of glycoGag on Ser5 endocytosis | 第35-36页 |
| 2.2.7 Western blotting | 第36页 |
| 2.2.8 Confocal microscopy | 第36-37页 |
| 2.2.9 Co-immunoprecipitation | 第37页 |
| 2.2.10 Flow cytometry | 第37页 |
| 2.2.11 Statistical analysis | 第37-38页 |
| 2.3 Results | 第38-57页 |
| 2.3.1 Plasmids confirmation | 第38页 |
| 2.3.2 Ser5 restricts MLV infectivity | 第38-39页 |
| 2.3.3 Ser5 incorporates into MLV particles | 第39-40页 |
| 2.3.4 GlycoMA and Nef counteracts Ser5 restriction and restore HIV-1 infectivity | 第40-41页 |
| 2.3.5 GlycoMA downregulates Ser5 expression | 第41-42页 |
| 2.3.6 GlycoMA re-localizes Ser5 to cytoplasmic compartments | 第42页 |
| 2.3.7 GlycoMA reduces Ser5 cell surface expression | 第42-43页 |
| 2.3.8 GlycoMA internalizes Ser5 via endocytosis | 第43-45页 |
| 2.3.9 GlycoMA utilizes Ap-2 pathway for Ser5 endocytosis | 第45页 |
| 2.3.10 Identification of crucial glycoGag residues responsible for Ser5 downregulation | 第45-48页 |
| 2.3.11 Intracellular interactions of glycoMA with Ser | 第48-50页 |
| 2.3.12 Rab GTPases are involved in Ser5 downregulation | 第50-52页 |
| 2.3.13 Ubiquitination plays crucial role in glycoMA-mediated Ser5 downregulation | 第52-54页 |
| 2.3.14 GlycoMA targets Ser5 to lysosomes for degradation | 第54-56页 |
| 2.3.15 GlycoMA downregulates murine Ser1,Ser2 and Ser | 第56-57页 |
| 2.4 Discussion | 第57-59页 |
| CHAPTER 3 MECHANISTIC INVESTIGATIONS OF SERINC5 COUNTERACTION TRIGGERED BY EQUINE INFECTIOUS ANEMIA VIRUS ACCESSORY PROTEIN S2 | 第59-83页 |
| 3.1 Background | 第59-60页 |
| 3.2 Material and methods | 第60-63页 |
| 3.2.1 Cells and culturing system | 第60页 |
| 3.2.2 Plasmids | 第60-61页 |
| 3.2.3 Transfection | 第61页 |
| 3.2.4 Effect of S2 on HIV-1?N infectivity in the presence mSer | 第61-62页 |
| 3.2.5 Effect of S2 on Ser5 endocytosis | 第62页 |
| 3.2.6 Western blotting | 第62页 |
| 3.2.7 Confocal microscopy | 第62页 |
| 3.2.8 Flow cytometry | 第62页 |
| 3.2.9 Co-Immunoprecipitation | 第62页 |
| 3.2.10 Statistical analysis | 第62-63页 |
| 3.3 Results | 第63-82页 |
| 3.3.1 Plasmids confirmation | 第63页 |
| 3.3.2 S2 rescues HIV-1 infectivity and counteract Ser5 restriction | 第63-64页 |
| 3.3.3 S2 triggered Ser5 down-regulation | 第64-65页 |
| 3.3.4 S2 internalizes Ser5 to cytoplasmic compartments | 第65-66页 |
| 3.3.5 Crucial residues of S2 for interaction with Ser | 第66-68页 |
| 3.3.6 Intracellular interactions of S2 and Ser | 第68-70页 |
| 3.3.7 S2 internalizes Ser5 via endocytosis | 第70-71页 |
| 3.3.8 S2 utilizes AP-2 pathway for Ser5 internalization | 第71-72页 |
| 3.3.9 Rab GTPases are involved in Ser5 downregulation | 第72-74页 |
| 3.3.10 Ubiquitination plays crucial role in S2-mediated Ser5 downregulation | 第74-75页 |
| 3.3.11 S2 targets Ser5 to lysosomes for degradation | 第75-76页 |
| 3.3.12 Equine Ser5 restricts retroviral infectivity and counteracted by S | 第76-78页 |
| 3.3.13 S2 downregulates other members of SERINC family(Ser1,Ser2 and Ser3) | 第78-79页 |
| 3.3.14 S2 mediated downregulation of Ser5 is stronger than Nef | 第79-80页 |
| 3.3.15 S2 and glycoMA are broader antagonists of Ser5 than Nef | 第80-82页 |
| 3.4 Discussion | 第82-83页 |
| CHAPTER 4 CONCLUSIONS | 第83-84页 |
| REFERENCES | 第84-95页 |
| Curriculum Vitae | 第95页 |
