Identification and characterization of ribonucleoproteins associated with genomic and antigenomic RNA during active EV71 replication

• Main Authors: Yu-Chun Chen, 陳郁淳
• Other Authors: Yeou-Guang Tsay
• Format: Others
• Language: en_US
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/24919199833548590984

碩士 === 國立陽明大學 === 生化暨分子生物研究所 === 101 === Enterovirus type 71(EV71) has emerged as an important non-polio neurological disease around Asia-Pacific regions, especially in Taiwan. EV71 cases with severe complication and outbreaks have been reported annually since 1998. Until now, there is no effective therapy against EV71, which partly results from lack of knowledge of EV71 replication. Here, we are interested in what host factors are assembled into viral replication protein complexes, more specifically, what host factors are closely associated with viral genome and antigenome. In this thesis, we first established a proteomic assay for isolation of ribonucleoproteins associated with specific RNA via UV irradiation. Coupled with two approaches to removal of non-specific signals, this platform totally gave 19 host factors associated with viral genomic RNA. In addition to abundant hnRNPs, these RNA-binding proteins include mitochondrial enzymes and structural proteins, suggesting the roles of energy suppliers and structural support in viral replication. Meanwhile, only one protein, desmin, was bound to antigenomic RNA, the reason of which might be low amount of these RNAs. In conclusion, our procedure can isolate ribonucleoproteins bound to EV71 genome and antigenome in vivo. While our procedure for specific elution should be further optimized, some of these results indeed shed light on future strategies on preparation and characterization of viral replication complexes.