热休克蛋白90(Hsp90)调控猪圆环病毒2型复制的分子机制

项目名称： 热休克蛋白90(Hsp90)调控猪圆环病毒2型复制的分子机制

项目编号： No.31502063

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： 刘捷

作者单位： 中国人民解放军东部战区总医院

项目金额： 21万元

中文摘要： 猪圆环病毒2型（PCV2）是一种严重危害猪体健康，对养猪业具有重要影响的病原，但其致病机制尚不十分清楚。热休克蛋白（HSPs）是一类高度保守的蛋白质，在细胞生长分化、基因转录、蛋白质合成-折叠-分解等方面发挥重要生理作用。调节Hsp90表达能够影响多种病毒在宿主细胞中的复制能力。我们前期研究发现，PCV2感染的PAMs和PK-15细胞中Hsp90表达水平明显升高。本研究拟通过Hsp90抑制剂处理细胞，利用真核质粒过表达和靶基因siRNA干扰技术调节细胞内Hsp90表达量，观察PCV2复制水平变化，明确Hsp90α/Hsp90β对PCV2复制的影响及其与PCV2 ORFs的相互作用关系；同时检测调节Hsp90表达影响PCV2复制的信号通路分子变化，包括Hsp90客户蛋白、NF-κB通路和蛋白酶体途径，探明Hsp90调控PCV2复制的分子机制，有利于阐明PCV2致病机制和提供新的防控策略。

中文关键词： 猪圆环病毒2型；热休克蛋白90；复制；分子机制

英文摘要： Porcine circovirus type 2 (PCV2) is one of the most important swine infectious diseases, which has spread worldwide. But its pathogenic mechanism remains poorly understood. Heat shock proteins (HSPs) are a group of highly conserved cellular proteins, which play important roles in cell growth and differentiation, gene transcription, protein synthesis, folding and degradation. Recently, Hsp90 was found to be an important host factor for the replication of some viruses. Our previous results showed that Hsp90 upregulated in PCV2 infected PAMs and PK-15 cells. In this study, we will detect the effect of inhibition of Hsp90 on PCV2 replication. The roles of Hsp90α or Hsp90β in PCV2 replication will be tested after down-regulation of Hsp90α/β via RNA interference and up-regulation of Hsp90α/β via transfection of recombinant plasmids. The interaction of Hsp90α/β with viral ORFs will be identified by confocal microscopy and Co-IP. Furthermore, the signal pathways of Hsp90 client proteins, NF-κB pathway and ubiquitin-proteasome system will be examined, respectively. These results will be helpful for understanding the mechanism of replication and pathogenesis of PCV2 and development of novel antiviral therapies in the future.

英文关键词： PCV2;Hsp90;replication;molecular mechanism