SERINC家族屬于多重跨膜蛋白,這個(gè)家族包括5個(gè)成員,但各成員的功能目前并不清楚。其中SERINC5（Ser5）作為逆轉(zhuǎn)錄病毒限制因子,通過未知機(jī)制插入到病毒粒子中并抑制病毒侵入下一個(gè)靶細(xì)胞。研究發(fā)現(xiàn)人免疫缺陷病毒（HIV-1）的附屬蛋白Nef能將Ser5內(nèi)化到細(xì)胞漿中并通過溶酶體降解Ser5,進(jìn)而阻止Ser5進(jìn)入病毒粒子。除了HIV-1 Nef蛋白之外,鼠白血病病毒（MLV）的附屬蛋白glycoGag以及馬傳染性貧血病毒（EIAV）的附屬蛋白S2也能拮抗Ser5對HIV-1病毒的限制作用。然而,MLV glycoGag以及EIAV S2拮抗Ser5的機(jī)制目前還不清楚。在本論文中,我們系統(tǒng)探討了glycoGag及S2如何拮抗Ser5蛋白的抗病毒活性。我們研究發(fā)現(xiàn)Ser5能插入到MLV的病毒粒子中并限制其感染性,而glycoGag能阻止Ser5插入到病毒粒子中并拮抗Ser5的抗病毒活性。研究發(fā)現(xiàn)glycoMA,既GlycoGag蛋白N端的189個(gè)氨基酸能發(fā)揮和glycoGag一樣的功能,因此我們利用glycoMA深入探討了其拮抗Ser5的詳細(xì)機(jī)制。我們發(fā)現(xiàn)glycoMA能將Ser5從細(xì)... 

【文章來源】：中國農(nóng)業(yè)科學(xué)院北京市

【文章頁數(shù)】：95 頁

【學(xué)位級別】：博士

【文章目錄】：
摘要
abstract
Abbreviations
CHAPTER 1 INTRODUCTION
    1.1 Restriction factors
        1.1.1 Discovery and molecular biology of SERINC genes
        1.1.2 Identification of Ser5 and Ser3 as restriction factors for HIV-
        1.1.3 Ser5 mediated inhibition of viral infectivity
        1.1.4 Mechanistic understanding of viral fusion inhibition triggered by Ser
    1.2 Retroviruses
        1.2.1 HIV-1
        1.2.2 MLV glycoGag
        1.2.3 EIAV S2
    1.3 Aims of the project
CHAPTER 2 MURINE LEUKEMIA VIRUS ACCESSORY PROTEINS GLYCOGAG REDUCES MURINE SERINC5 PROTEIN EXPRESSION LEVEL VIA DEGRADING LYSOSOMAL PATHWAY TO COUNTERACT SERINC5 ANTIRETROVIRAL ACTIVITY
    2.1 Background
    2.2 Material and methods
        2.2.1 Cells and culturing system
        2.2.2 Plasmids
        2.2.3 Transfection
        2.2.4 Effect of mSer5 on HIV-1 infectivity
        2.2.5 Effect of mSer5 on MLV infectivity
        2.2.6 Effect of glycoGag on Ser5 endocytosis
        2.2.7 Western blotting
        2.2.8 Confocal microscopy
        2.2.9 Co-immunoprecipitation
        2.2.10 Flow cytometry
        2.2.11 Statistical analysis
    2.3 Results
        2.3.1 Plasmids confirmation
        2.3.2 Ser5 restricts MLV infectivity
        2.3.3 Ser5 incorporates into MLV particles
        2.3.4 GlycoMA and Nef counteracts Ser5 restriction and restore HIV-1 infectivity
        2.3.5 GlycoMA downregulates Ser5 expression
        2.3.6 GlycoMA re-localizes Ser5 to cytoplasmic compartments
        2.3.7 GlycoMA reduces Ser5 cell surface expression
        2.3.8 GlycoMA internalizes Ser5 via endocytosis
        2.3.9 GlycoMA utilizes Ap-2 pathway for Ser5 endocytosis
        2.3.10 Identification of crucial glycoGag residues responsible for Ser5 downregulation
        2.3.11 Intracellular interactions of glycoMA with Ser
        2.3.12 Rab GTPases are involved in Ser5 downregulation
        2.3.13 Ubiquitination plays crucial role in glycoMA-mediated Ser5 downregulation
        2.3.14 GlycoMA targets Ser5 to lysosomes for degradation
        2.3.15 GlycoMA downregulates murine Ser1,Ser2 and Ser
    2.4 Discussion
CHAPTER 3 MECHANISTIC INVESTIGATIONS OF SERINC5 COUNTERACTION TRIGGERED BY EQUINE INFECTIOUS ANEMIA VIRUS ACCESSORY PROTEIN S2
    3.1 Background
    3.2 Material and methods
        3.2.1 Cells and culturing system
        3.2.2 Plasmids
        3.2.3 Transfection
        3.2.4 Effect of S2 on HIV-1?N infectivity in the presence mSer
        3.2.5 Effect of S2 on Ser5 endocytosis
        3.2.6 Western blotting
        3.2.7 Confocal microscopy
        3.2.8 Flow cytometry
        3.2.9 Co-Immunoprecipitation
        3.2.10 Statistical analysis
    3.3 Results
        3.3.1 Plasmids confirmation
        3.3.2 S2 rescues HIV-1 infectivity and counteract Ser5 restriction
        3.3.3 S2 triggered Ser5 down-regulation
        3.3.4 S2 internalizes Ser5 to cytoplasmic compartments
        3.3.5 Crucial residues of S2 for interaction with Ser
        3.3.6 Intracellular interactions of S2 and Ser
        3.3.7 S2 internalizes Ser5 via endocytosis
        3.3.8 S2 utilizes AP-2 pathway for Ser5 internalization
        3.3.9 Rab GTPases are involved in Ser5 downregulation
        3.3.10 Ubiquitination plays crucial role in S2-mediated Ser5 downregulation
        3.3.11 S2 targets Ser5 to lysosomes for degradation
        3.3.12 Equine Ser5 restricts retroviral infectivity and counteracted by S
        3.3.13 S2 downregulates other members of SERINC family（Ser1,Ser2 and Ser3）
        3.3.14 S2 mediated downregulation of Ser5 is stronger than Nef
        3.3.15 S2 and glycoMA are broader antagonists of Ser5 than Nef
    3.4 Discussion
CHAPTER 4 CONCLUSIONS
REFERENCES
Curriculum Vitae



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