HSP90AB1与IBV S1蛋白互作调节病毒吸附的研究  

Study on the effects of HSP90AB1 interacting with IBV S1 protein on virus attachment

作　　者：张焕东 方承秀 林翠 董伟仁[1] 周继勇[1] 廖敏[1] ZHANG Huan-dong;FANG Cheng-xiu;LIN Cui;DONG Wei-ren;ZHOU Ji-yong;LIAO Min(Key Laboratory of Animal Virology of Ministry of Agriculture and Rural Affairs/Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学农业农村部动物病毒学重点实验室,浙江杭州310058

出　　处：《中国兽医科学》2023年第8期983-990,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2021YFD1801102)。

摘　　要：前期的免疫沉淀和质谱鉴定分析表明,HSP90AB1是禽传染性支气管炎病毒(IBV)S1蛋白的潜在互作宿主蛋白。本研究在此基础上探讨HSP90AB1与IBV S1蛋白的相互作用及其对IBV吸附宿主细胞的影响。免疫共沉淀试验显示,宿主HSP90AB1蛋白可与多种IBV毒株的S1蛋白发生互作,Gst-pull-down试验证明,HSP90AB1与M41株S1蛋白的互作属于直接互作。进一步的免疫沉淀试验表明,细胞膜HSP90AB1与内源性S1蛋白同样具有互作关系。进一步研究发现过表达HSP90AB1会促进IBV Beaudette对Vero细胞的吸附,而敲低HSP90AB1则抑制病毒吸附H1299细胞;使用商品化的HSP90AB1家兔单克隆抗体预处理Vero细胞则抑制了病毒的吸附;用HSP90AB1蛋白阻断结合位点使病毒吸附细胞的能力显著降低。以上结果表明,HSP90AB1对IBV吸附细胞具有正调节作用,其可能是IBV入侵细胞的一个关键宿主因子。Previous immunoprecipitation and mass spectrometry analysis showed that HSP90AB1 was a potential host protein interacting with S1 protein of avian infectious bronchitis virus(IBV).On this basis,this study explored the interaction between HSP90AB1 and IBV S1 protein and its effects on the attachment of IBV to host cells.Immunocoprecipitation test showed that the host HSP90AB1 protein could interact with S1 protein of different genotype IBV strains,and Gst-pull down test showed that the S1 protein of M41 strain directly bind to HSP90AB1.Further immunoprecipitation test showed that the cell membrane HSP90AB1 also interacted with endogenous S1 protein.Further study showed that overexpression of HSP90AB1 promoted the attachment of IBV Beaudette on Vero cells,while knockdown of HSP90AB1 inhibited the attachment of virus on H1299 cells;Pretreatment of Vero cells with commercialized rabbit monoclonal antibody to HSP90AB1 inhibited virus attachment;Blocking the binding site with HSP90AB1 protein significantly reduced the attachment of the virus to host cells.The above findings indicate that HSP90AB1 has a positive regulatory effects on IBV attachment,which may be a key host factor for IBV invasion to host cells.

关 键 词：禽传染性支气管炎病毒 热休克蛋白HSP90AB1 S1蛋白 病毒吸附 

分 类 号：S852.659.6[农业科学—基础兽医学]