The Functional Studies of Enterovirus 71 2C Protein in Membrane Binding and Rearrangement

• Main Authors: Wen-fang Tang, 湯文芳
• Other Authors: Jim-tong Horng
• Format: Others
• Language: zh-TW
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/79616518411074252169

碩士 === 長庚大學 === 基礎醫學研究所 === 92 === Enterovirus 71 (EV71) is an enterovirus of the family Picornaviridae. In 1998, an EV71 epidemic in Taiwan was associated with hand, foot, and mouth disease (HFMD)/herpangina. Currently, there is no vaccine available for non-polio enteroviruses. Previous studies show that poliovirus (PV) 2C protein was essential for virus replication. In PV-infected cells, 2C protein is associated membrane vesicles that surround the viral replication complexes. Recombinant PV 2C protein expressed in transfected cells has been shown to associate with and induce rearrangement of the intracellular membrane network. However, the 2C protein is highly conserved viral protein among all picornaviruses. We are interested to know what proteins might interact with EV71 2C protein during virus replication. We observed that N-terminus of EV71 2C protein can interact with reticulon 1-C (RTN1C) and reticulon 3 (RTN3) by yeast two-hybrid screen. We further confirmed that the interaction is genuine and specific by co-immunoprecipitation and by immunofluorescence microscopy. The interaction region of RTN3 was mapped to amino acids 69-168 and that of 2C was amino acids 5-43 by co-immunoprecipitation assay. By sucrose gradient ultracentrifugation, RTN3 was found to associate 2C and 3A which are involved in virus replication. This suggested that RTN3 might play a role in replication. RTN3 can also interact with PV2C, implying that RTN3 is a common factor for both EV71 and PV replication. Direct evidence for the function of RTN3 is underway by siRNA assay.