Real-time visualization of intracellular infection route of Dengue virus
| Autor: | Yi-Lung Huang, 黃俋龍 |
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| Rok vydání: | 2011 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 99 Understanding of the molecular mechanisms of the cellular trafficking pathway of virus in living cells is one of the major goals in modern virology. The real-time single particle tracking technique allows us to comprehend the dynamics of interaction between virus and host cells. Dengue virus (DENV) is the enveloped, positive-sense single-stranded RNA virus that causes the most common arthropod-borne infective disease globally. Previous studies have reported that DENV utilizes clathrin-mediated endocytosis to enter host cell. However, the intracellular pathways of DENV after entering cells remain to be uncovered. To address this question, we investigated the intracellular pathway of DENV by tracking a single viral particle labeled with a lipophilic fluorescent dye, DiD, in living cells that express fluorescent markers of cellular compartments. The intracellular trajectory of each single DENV particle was recorded by a home-built real-time multi-color fluorescence microscopy. By using chlorpromazine (CPZ) to block clathrin pits formation, we found that 80 % of DENV particles entered cells majority through clathrin-mediated endocytosis. Next, we tracked DiD-labeled DENV particles in living cells expressing LC3 fused GFP (GFP-LC3) for exploring the itinerary of cellular compartments. Following clathrin-mediated uptake, the majority of DENV particles were transported to LC3-positive autophagosomes where membrane fusion could be detected as the disappearance of DiD signals. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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