Investigation of the intracellular trafficking pathway of vaccinia virus in HeLa cells

• Main Authors: Tzu-Jung Chen, 陳姿融
• Other Authors: Wen Chang
• Format: Others
• Language: en_US
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/51213580262036727129

碩士 === 國立陽明大學 === 生命科學暨基因體科學研究所 === 99 === Vaccinia virus is the prototypic member of the Orthopoxvirus genus and has a wide host range in vitro and in vivo. Vaccinia virus contains an envelope and enters cells through either endocytosis or plasma membrane fusion. We previously showed that the WR strain of vaccinia virus entered HeLa cells through fluid-phase endocytosis process that depends on cellular proteins dynamin and vaccinia virus penentration factor (VPEF) but not clathrin nor caveolae. However, after being uptake into cells, the intracellular trafficking route of the internalized vaccinia virus remains unclear. The goal of my thesis is to elucidate the trafficking process of internalized vaccinia mature virus inside the cells until membrane fusion that releases viral cores in cytoplasm. Here, we utilized two sets of experimental designs to address this issue. First we monitored mCherry-labeled virus particles uptake process in order to identify virus transport from PI3P-enriched macropinosomes to Rab5+endosome and other intracellular compartments prior to membrane fusion. Furthermore, using functional interference assays, we perturbed different intracellular vesicle trafficking processes in order to delineate the intracellular trafficking pathway of the internalized virions prior to membrane fusion. The results showed that vaccinia virus was internalized and transported to Rab5+ early endosomes but not Rab7+ late endosomes. Instead, Rab11+ recycling endosomes appeared involved in virus trafficking in cells. Future work will be performed to verify the role of retromer complexes in virus endocytic entry process and to investigate how rab11 and retromer complex interact to regulate vaccinia virus trafficking.