动脉炎病毒属的RNA解旋酶的结构和功能研究

Arteritis viruses, such as porcine reproductive and respiratory syndrome virus (PRRSV) and equine arteritis virus (EAV), are positive-strand RNA viruses and seriously harm the livestock and cause significant economic loss. The RNA viruses varied rapidly during replication, making the development of viral vaccines very difficult. To date, no effective method for the treatment and prevention of the viruses was found. It was found that the RNA helicases (nsp10) were among the most conserved proteins in the arterivirus proteins, played very important roles in various cellular processes. In addition, the nsp10 family of arterivirus shows high homology with the RNA helicase from the coronaviruses which greatly hurt human health. The nsp10 helicase contains a zinc binding domain, and a core helicase domain which can bind nucleoside triphosphate and RNA. Currently, no structures of nsp10 or homology have been solved. Based on present work, this study would focus on solving the structures of the nsp10 family, and complex structures with nucleoside triphosphate and DNA/RNA by protein crystallography. The project will uncover the regulatory mechanism between domains of nsp10, substrate selectivity and specificity, the relationship of structure and function, and the mechanism of catalysis and unwinding. It will provide a structural basis and a direction for the search of interacting proteins, screening inhibitors and drug design.

动脉炎病毒属是正链RNA病毒，常见的有猪繁殖与呼吸综合征病毒(PRRSV)和马动脉炎病毒(EAV)，给畜牧业造成重大的危害和经济损失。病毒在复制过程中会快速变异，造成病毒疫苗的研制步履维艰，目前还没有特别有效的方法来治疗和预防。实验发现核酸解旋酶(nsp10)是此类病毒蛋白中最保守的一个蛋白，对病毒的多种细胞过程非常重要。另外，nsp10与对人类健康危害很大的冠状病毒的核酸解旋酶同源性很高。 nsp10含有一个锌离子结合结构域及一个能结合RNA和核苷三磷酸的解旋酶核心结构域。目前，nsp10或同源物的结构还没有被解析。本项目将从蛋白质晶体学角度解析nsp10家族及其与核苷三磷酸、DNA/RNA复合物的结构，研究nsp10内部结构域之间的调节机制、底物的选择性和特异性及其结构与功能的关系，深入探讨nsp10的催化和解旋机理。为寻找相互作用的蛋白、筛选抑制剂和药物设计等提供结构基础和指明方向。

动脉炎病毒属是正链RNA病毒，常见的有猪繁殖与呼吸综合征病毒(PRRSV)和马动脉炎病毒(EAV)，给畜牧业造成重大的危害和经济损失。病毒在复制过程中会快速变异，造成病毒疫苗的研制步履维艰，目前还没有特别有效的方法来治疗和预防。实验发现核酸解旋酶(nsp10)是此类病毒蛋白中最保守的一个蛋白，对病毒的多种细胞过程非常重要。另外，nsp10与对人类健康危害很大的冠状病毒的核酸解旋酶同源性很高。.在本基金的资助下，围绕项目的计划，本项目在PRRSV和EAV的nsp10结构及其与核酸复合物结构的解析与功能研究方面，取得一些富有意义的成果。培养高素质的研究生13人。标有本课题资助、以通信作者或共同通信作者发表的SCI论文有12篇,发表在有国际影响力的重要期刊如：Genes & Development 、Nature Communications 、PNAS 、Nucleic Acids Research 、JBC和J. Virol等杂志上。首次解析出动脉炎病毒属解旋酶及其复合物结构，发现马动脉炎病毒属解旋酶三维结构保守，其N端的ZBD结构域采用一种全新的折叠方式。EAV nsp10的酶活力依赖于ZBD和解旋酶核心区域HEL1之间的互作。为这个重要的病毒家族蛋白寻找相互作用的蛋白、筛选抑制剂和药物设计等提供结构基础和指明方向。