| 猪丁型冠状病毒S1-CTD相互作用宿主蛋白的筛选和鉴定 |
| |
| 引用本文： | 汤荣锋,范前进,郭龙军,张鑫,石达,时洪艳,陈建飞,冯力.猪丁型冠状病毒S1-CTD相互作用宿主蛋白的筛选和鉴定[J].畜牧兽医学报,2022,53(7):2260-2267. |
| |
| 作者姓名： | 汤荣锋 范前进 郭龙军 张鑫 石达 时洪艳 陈建飞 冯力 |
| |
| 作者单位： | 中国农业科学院哈尔滨兽医研究所, 兽医生物技术国家重点实验室/猪消化道传染病创新团队, 哈尔滨 150069 |
| |
| 基金项目： | 兽医生物技术国家重点实验室自主研究课题(SKLVBP202105)； |
| |
| 摘 要： | 猪丁型冠状病毒纤突蛋白S1亚基C端结合域(S1-CTD)是诱导中和抗体产生的主要区域，为研究与其相互作用的宿主蛋白，采用HEK-293T真核表达系统表达并纯化了S1-CTD，提取了猪回肠上皮细胞膜蛋白，通过免疫共沉淀筛选了可能与S1-CTD相互作用的蛋白并进行质谱分析，发现32个疑似相互作用的宿主蛋白。构建其中可能与S1-CTD互作的蛋白KIF1 binding protein (KIFBP)的真核表达质粒，通过免疫共沉淀和激光共聚焦验证上述宿主蛋白与S1-CTD是否存在相互作用，结果表明KIFBP和S1-CTD之间存在相互作用，共同转染时在细胞质共定位。进一步研究表明，过表达KIFBP能够有效降低病毒RNA水平和病毒滴度，其中mRNA水平降低了约70%，病毒滴度降低了101.6TCID50。综上，本研究筛选并鉴定出一种与PDCoV S1-CTD相互作用的宿主蛋白KIFBP，为了解PDCoV的致病机制提供了理论基础。
|
| 关 键 词： | 猪丁型冠状病毒 S1-CTD蛋白 相互作用 KIFBP |
| 收稿时间： | 2021-10-26 |
Screening and Identification of Host Proteins Interacting with PDCoV S1-CTD |
| |
| TANG Rongfeng,FAN Qianjin,GUO Longjun,ZHANG Xin,SHI Da,SHI Hongyan,CHEN Jianfei,FENG Li.Screening and Identification of Host Proteins Interacting with PDCoV S1-CTD[J].Acta Veterinaria et Zootechnica Sinica,2022,53(7):2260-2267. |
| |
| Authors: | TANG Rongfeng FAN Qianjin GUO Longjun ZHANG Xin SHI Da SHI Hongyan CHEN Jianfei FENG Li |
| |
| Institution: | Swine Digestive System Infectious Diseases Division, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, State Key Laboratory of Veterinary Biotechnology, Harbin 150069, China |
| |
| Abstract: | The C-terminal domain (CTD) of porcine deltacoronavirus S1 subunit is the main region which induces the neutralizing antibody. S1-CTD was expressed by HEK-293T eukaryotic expression system and purified, and porcine ileal epithelium cells membrane proteins were extracted to investigate porcine host proteins that interact with it. Thirty-two suspected interacting host proteins were obtained by co-inmunprecipitation (Co-IP) and mass spectrometry. Eukaryotic expression plasmid of KIF1 binding protein (KIFBP) was constructed, and the interaction between KIFBP and S1-CTD was identified by Co-IP and laser confocal microscopy. All results proved that KIFBP interacted with S1-CTD and co-located in cytoplasm. Further research indicated that overexpression of KIFBP could effectively reduce the viral mRNA level and the viral titer in which the mRNA level decreased by about 70%, and the viral titer decreased by 101.6TCID50. In conclusion, a host protein KIFBP interacting with PDCoV S1-CTD was screened and identified in this study which provides a theoretical basis for understanding the pathogenesis of PDCoV. |
| |
| Keywords: | porcine deltacoronavirus S1-CTD protein interaction KIF1 binding protein |
|
| 点击此处可从《畜牧兽医学报》浏览原始摘要信息 |
| 点击此处可从《畜牧兽医学报》下载免费的PDF全文 |
