猪繁殖与呼吸综合征病毒N蛋白与宿主细胞DDX5蛋白相互作用研究  

CHARACTERIZATION OF INTERACTION BETWEEN NUCLEOCAPSID PROTEIN OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS AND HOST DDX5 PROTEIN

作　　者：刘欢[1] 黄勤锋[1] 张文超[1] 姜一峰[1] 杨莘[1] 高飞[1] 周艳君[1] 童光志[1] LIU Huan HUANG Qin-feng ZHANG Wen-chao JIANG Yi-feng YANG Shen GAO Fei ZHOU Yan-jun TONG Guang-zhi(Shanghai Veterinary Research Institute, CAAS, Shanghai 200241, China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2017年第3期12-17,共6页Chinese Journal of Animal Infectious Diseases

基　　金：国家科技支撑项目(2015BAD12B01-1);国家自然科学基金项目(31502072;31302098;31300140)

摘　　要：研究利用免疫共沉淀的方法证明猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)N蛋白与MARC-145细胞DDX5蛋白之间存在相互作用,且两种蛋白共定位于MARC-145细胞质内。利用构建的EGFP-N和m Cherry-DDX5真核表达载体共转染MARC-145细胞后,发现N蛋白并不能将DDX5蛋白募集至细胞质。采用si RNA沉默MARC-145细胞DDX5基因表达之后,病毒复制水平上升;荧光定量检测结果显示,DDX5沉默表达也能够明显促进病毒基因组RNA和长链亚基因组RNA的转录水平。结果表明DDX5参与了病毒复制转录过程并对病毒增殖具有抑制作用。In the present study, the interaction between Porcine reproductive and respiratory syndrome virus（PRRSV） N protein and cellular DDX5 protein was verified by co-immunoprecipitation assay. Furthermore, confocal laser experiment showed that PRRSV N protein and DDX5 colocalized in the cytoplasm of virus infected MARC-145 cells. In addition, eukaryotic vectors expressing EGFP-N and mCherry-DDX5 fusion proteins were transfected to MARC-145 cells. The results showed that DDX5 was not recruited into cytoplasm by N protein. When DDX5 was silenced by siRNA, the growth ability of PRRSV was enhanced and real-time RT-PCR results showed that the transcriptional level of genome RNA and longer subgenomic RNA was significantly higher than that of the control, which indicating a negative regulatory role of DDX5 during PRRSV replication and transcription.

关 键 词：猪繁殖与呼吸综合征病毒 N蛋白 DDX5 相互作用 

分 类 号：S852.659.6[农业科学—基础兽医学]