FUNCTIONAL ANALYSIS OF DENGUE VIRUS ANTIGENS NS3 AND NS5

描述：提案的总体目标是阐明 登革病毒2型NS 3和NS 5蛋白的多重功能 在病毒RNA复制和5'基因组RNA加帽中。 一个明确 了解病毒RNA复制对于了解 病毒致病机制。NS 3与NS 2B联合 蛋白质是加工病毒所需的丝氨酸蛋白酶， 多聚蛋白 NS 3还具有NTR/RNA的保守基序 解旋酶，并且它还具有5' RNA三磷酸酶活性，这是 病毒复制和加帽。 NS 5具有RNA基序 甲基转移酶和RNA依赖性RNA聚合酶。 体外RNA 将使用分离的膜结合复制来寻求合成 特异性依赖于外源添加的病毒RNA的复合物 含有基因组RNA的3'末端的模板。 病毒蛋白 负链合成所需的模板序列将 通过诱变鉴定。 NS 3的RNA解旋酶和RdRp 将表征参与复制的NS 5的活性。 在 此外，病毒蛋白及其酶活性所需的 将鉴定5' RNA加帽。 由于这些酶的活性是 它们是病毒生命周期中唯一需要的，是潜在的目标 用于抗病毒策略。

DESCRIPTION: The overall aim of the proposal is to elucidate the multiple functions of the NS3 and NS5 proteins of dengue virus type 2 in viral RNA replication and 5' genomic RNA capping. A clear understanding of viral RNA replication is important to understand the mechanism of viral pathogenesis. NS3 in association with the NS2B protein is a serine protease required for the processing of the viral polyprotein. NS3 also has the conserved motifs of a NTPase/RNA helicase, and it also has a 5' RNA triphosphatase activity, required for viral replication and capping respectively. NS5 has motifs for RNA methyltransferase and RNA-dependent RNA polymerase. In vitro RNA synthesis will be sought for using isolated membrane bound replication complexes that are specifically dependent on exogenously added viral RNA templates containing the 3' terminus of the genomic RNA. Viral proteins and template sequences required for the minus strand synthesis will be identified by mutagenesis. The RNA helicase of NS3 and the RdRp activity of NS5 involved in replication will be characterized. In addition, the viral proteins and their enzyme activities required for 5' RNA capping will be identified. Since these enzyme activities are uniquely required for the virus lifecycle, they are potential targets for antiviral strategies.