Mechanistic study for the interaction between cellular factor and hepatitis C virus RNA polymerase NS5B

• Main Authors: Hung-Ju Chiang, 姜宏儒
• Other Authors: Ju-Chien Cheng
• Format: Others
• Language: en_US
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/44j49n

碩士 === 中國醫藥大學 === 醫學檢驗生物技術學系碩士班 === 102 === Hepatitis C virus (HCV) is one of the major pathogens that cause liver diseases. The viral nonstructural protein 5B (NS5B) is a RNA-dependent RNA polymerase (RdRp) that plays the major role in HCV replication. It has been discovered that NS5B interacts with various host factors in order to alter host physiological activities. Previous studies in our laboratory demonstrated that nuclear receptor Nur77 interacted with HCV NS5B by yeast two-hybrid (Y2H) analysis. To confirm the interaction between Nur77 and NS5B, GST pull down assay and immunoprecipitation assay were performed. The results indicated that Nur77 interacts with HCV NS5B both in vitro and in vivo. Immunofluorescence assay was also operated to display the sub-cellular localization of the interaction between Nur77 and NS5B. Moreover, the ratio of Nur77 nucleus exportation was observed under the presence of NS5B. It has been reported that phosphorylation status regulates Nur77 subcellular trafficking. A mobility shift of NS5B-interacting Nur77 was observed and the increased mass of Nur77 might be through NS5B induced kinase activity. Taken together, our results suggest that NS5B facilitates Nur77 shuttling from nuclear to cytoplasm to interact with each other. Further investigations focusing on NS5B involved in Nur77-mediated biological functions are required to be answered.