Mechanisms by Which Enterovirus-Host Interactions Form Viral Replication Organelles and Regulate Viral Replication

All positive-strand RNA viruses recognize intracellular membranes in replication organelles to amplify their genomes. However, various viruses hijack host factors and membranes of various organelles. For instance, flaviviruses develop a membrane web from the endoplasmic reticulum (ER), while enteroviruses form new tubular-vesicular organelles using a Golgi membrane. Picornaviruses are a diverse and major cause of human disease, and their genomes replicate with intracellular membranes. The functionality of these replication organelles depends on the activities of both the viral nonstructural proteins and the co-opted host proteins. The mechanism by which viral-host interactions generate viral replication organelles and regulate viral RNA synthesis is unclear. To elucidate this mechanism, enterovirus 71 (EV71) is used herein as a virus model to investigate how these replication organelles are formed and to identify the cellular components that are critical in this process. Immunoprecipitation assay is combined with LC-MS/MS analysis to identify 172 cellular proteins and four viral proteins. SCAMP3 is one of the host proteins that is selected for further investigation. This project has the following goals. Aim I. To investigate the interaction of viral proteins with SCAMP3 Aim II. To investigate the role of SCAMP3 in viral replication Aim III. To identify whether SCAMP3 involved another pathway for replication organelles and the role of SCAMP3 in other enteroviruses replication Understanding how viral proteins hijack the regulatory mechanisms of membrane metabolism to form replication organelles will provide a new perspective on various areas of cell biology and virology and will be indispensable for the development of a new generation of anti-viral control strategies.