Takegami T, Sakamuro D, Furukawa T
Division of Tropical Medicine, Kanazawa Medical University, Ishikawa, Japan.
Virus Genes. 1995 Jan;9(2):105-12. doi: 10.1007/BF01702653.
Sequence data suggest that Japanese encephalitis virus (JEV) protein NS3 is a multifunctional protein with sequence motifs characteristic of a protease and a helicase. To examine the functions of JEV-NS3, a fusion protein of NS3 in Escherichia coli was generated. Analysis by Western blot using monospecific rabbit antisera generated against the fusion protein (anti-MBJEN3) showed that NS3 was localized in the membrane fraction of JEV-infected cells and the particulate fraction of bacteria extracts. The addition of anti-MBJEN3 sera reduced JEV-specific RNA synthesis activity in a in vitro system. In addition, NS3 was shown to exhibit RNA binding and ATPase activities, suggesting this protein has an important role in viral RNA replication in virus-infected cells.
序列数据表明，日本脑炎病毒（JEV）的NS3蛋白是一种多功能蛋白，具有蛋白酶和解旋酶的序列基序特征。为了研究JEV-NS3的功能，在大肠杆菌中构建了NS3融合蛋白。使用针对该融合蛋白产生的单特异性兔抗血清（抗MBJEN3）进行的蛋白质印迹分析表明，NS3定位于JEV感染细胞的膜组分和细菌提取物的颗粒组分中。在体外系统中加入抗MBJEN3血清可降低JEV特异性RNA合成活性。此外，NS3显示出具有RNA结合和ATP酶活性，表明该蛋白在病毒感染细胞的病毒RNA复制中起重要作用。
