发热伴血小板减少综合征病毒核蛋白特异结合宿主细胞60kD SSA/Ro

Severe Fever with Thrombocytopenia Syndrome Virus Nucleoprotein Specifically Binds to 60kD SSA/Ro Protein in Host Cells

为了初步探索发热伴血小板减少综合征病毒(Severe fever with thrombocytopenia syndrome virus,SFTSV)核蛋白(nucleoprotein,NP)对宿主细胞免疫功能的影响。将SFTSV NP和NSs蛋白的编码基因插入真核表达载体VR1012中,通过免疫共沉淀(IP)、SDS-PAGE、质谱检测及蛋白质免疫印迹等方法寻找宿主细胞中与NP相互作用,同时又与免疫功能有关的蛋白质分子,并利用细胞免疫荧光方法检测SFTSV NP与该分子在细胞中的共定位情况。IP和质谱检测结果显示NP能与免疫功能相关的60kD SSA/Ro蛋白发生特异性结合,细胞免疫荧光试验进一步显示NP与60kD SSA/Ro蛋白在细胞质中存在共定位。提示SFTSV可能通过其核蛋白与免疫相关分子60kD SSA/Ro蛋白发生特异性结合,从而引起机体一系列的免疫反应和临床症状。

This study aims to investigate whether the nucleoprotein （NP） of severe fever with thrombocytopenia syndrome virus （SFTSV） can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein （NSs） of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation （IP）, SDS-PAGE, mass spectrometry （MS）, and Western blot. Co-localization of NP and the identified host proteins was confirmed by eonfocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confoeal microscopy showed that NP and SSA/Ro were colocalized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.