| Summary: | 碩士 === 臺北醫學大學 === 細胞及分子生物研究所 === 93 === Vaccinia virus (VV) is a poxvirus that replicates in cytoplasm of the host cells. The DNA genome of vv is 192 Kb in size and encodes 263 viral proteins. In most cells, VV infection leads to production of virus progenies; however, in few cell types, virus life cycle was blocked at post-entry step, indicating the presence of host restriction in specific cell types. In one of such restricting cell line, CHO, VV replication requires a host range (hr) gene CP77 to complete life cycle. My thesis focuses on understanding the role of CP77 in host restriction regulation. The first part of my project is to investigate the structural and functional relationship of CP77. We construct recombinant VV expressing different CP77 deletions, monitor deletion protein expression and measure protein stability in HeLa cells and CHO cells. The results showed that the N-and C-terminal regions have protection effect, maintaining CP77 stability in virus-infected cells. The second part is to investigate the post-translational modifications on CP77. We found that CP77 is ubiquitinated in both HeLa and CHO virus-infected cells. Besides, we also showed that CP77 sumoylated in vitro. Finally, the third part, our previous data indicated that a cellular protein cytidine deaminase (CDA) interacts with CP77 in yeast two-hybrid analysis. I have showed that indeed CP77 and CDA interact in GST-pull down experiments, indicating a role of CDA in vaccinia virus host restriction regulation.
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