NSP5蛋白与宿主细胞beta tubulin相互作用对PoRV致病性的影响

项目名称： NSP5蛋白与宿主细胞beta tubulin相互作用对PoRV致病性的影响

项目编号： No.31502092

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 水产学、兽医学

项目作者： 时洪艳

作者单位： 中国农业科学院哈尔滨兽医研究所

项目金额： 18万元

中文摘要： 轮状病毒(Rotavirus，RV)是引起婴幼儿和多种幼龄动物病毒性腹泻的主要病原体。虽然关于RV的研究日益深入，但RV致病机理的许多方面仍然不清楚。猪轮状病毒(PoRV) NSP5是一个高度磷酸化的蛋白，在病毒颗粒装配中起到关键作用。本项目拟在发现PoRV NMTL株NSP5蛋白与beta tubulin相互作用的基础上，1）通过上调和下调beta tubulin表达，分析beta tubulin对PoRV在宿主细胞内复制的影响；2）通过对NSP5蛋白截短表达及定点突变，寻找NSP5蛋白与beta tubulin相互作用位点；3）通过反向遗传学技术拯救NSP5与beta tubulin相互作用区域缺失或突变的重组病毒，对重组病毒的生物学特性进行分析，以揭示NSP5与beta tubulin相互作用对PoRV的复制的影响，进而为深入了解PoRV致病机制提供新途径。

中文关键词： 猪轮状病毒；非结构蛋白5；微管蛋白；相互作用

英文摘要： Rotavirus (Rotavirus, RV) is the major pathogen causing diarrhea in infants and piglets. Although the research on RV is increasing, many aspects of the pathogenic mechanism of RV is still unclear. PoRV non-structural protein 5 (NSP5) is a highly phosphorylated protein, has RNA binding activity, plays a key role in the assembly of virus particles. Study on the interaction of NSP5 protein and the host cell, can deepen understanding of the pathogenic mechanism of PoRV. Interaction between PoRV NMTL strain NSP5 protein and beta tubulin was found. Based on the interaction, we want to 1) down-regulation or up re-gulating the expression of beta tubulin and analysis of the impact on PoRV replication in host cells; 2) through expression the truncated NSP5 protein and site directed mutagenesis, finding the interaction can site between NSP5 protein and beta tubulin; 3) rescue the virus deletion or mutation of NSP5 and beta tubulin interaction region to analysis of biological characteristics of the recombinant virus. The aim of this research is to reveal the NSP5 and beta tubulin interaction influence on PoRV replication, which provides a new way for the pathogenic mechanism of PoRV.

英文关键词： Porcine rotavirus;NSP5;Beta tubulin;Interaction