农业生物技术学报2024，Vol.32Issue(11)：2662-2669,8.DOI:10.3969/j.issn.1674-7968.2024.11.019
宿主蛋白SPCS1通过外泌体途径介导BVDV复制的研究
Research on Replication of BVDV Through Exosome Pathway Mediated by Host Protein SPCS1
摘要
Abstract
Bovine viral diarrhea(BVD)is an infectious disease of cattle worldwide caused by Bovine viral diarrhea virus(BVDV),which has brought huge economic losses to animal husbandry.The previous study found that knocked down the expression of signal peptidase complex subunit 1(SPCS1)in bovine kidney cells(Madin-Darby Bovine Kidney,MDBK)could inhibit the replication of BVDV effectively.In order to explore its molecular mechanism,this study collected exosomes from the supernatant of SPCS1 knockdown(KD)cells infected with BVDV,and transmission electron microscopy,Western blot and Nanoparticle Tracking Analysis were used to characterize exosomes.qPCR was used to analysis BVDV 5'untranslated region(UTR)RNA levels in the exosomes.The exosomes were infected with MDBK;BVDV 5'UTR RNA levels were detected by qPCR;The cytopathic effect(CPE)was observed by inverted microscope,and the change of virus titer was determined by Reed-Muench method.The exosomes were observed as disk-like vesicles under transmission electron microscopy,Western blot detected exosomes,the volume characteristic protein TSG101 showed a peak value at about 100 nm,indicating exosomes were successfully extracted.Western blot showed that the expression of exosome TSG101 protein in SPCS1 KD cells infected with BVDV was significantly decreased compared with the control cells treated with empty vector(Scramble cells)(P＜0.001).qPCR detection showed that the level of BVDV 5'UTR mRNA in exosomes was significantly decreased(P＜0.001).BVDV 5'UTR RNA levels of SPCS1 KD cell derived exosomes infected with MDBK were significantly lower than those infected with Scramble cell(P＜0.001),the CPE phenomenon was delayed and weakened,and the viral titer of the progeny was significantly decreased(P＜0.001).The results showed that knockdown SPCS1 affected the replication of BVDV through exosome pathway.The research results provide a new antiviral target and theoretical basis for the prevention and control of BVDV.关键词
信号肽酶复合体亚基1(SPCS1)/牛病毒性腹泻病毒(BVDV)/外泌体Key words
Signal peptidase complex subunit 1(SPCS1)/Bovine viral diarrhea virus(BVDV)/Exosome分类
农业科技引用本文复制引用
张成远,魏玉荣,陈俊贞,杨莉,李泽宇,李紫仟,付强,史慧君..宿主蛋白SPCS1通过外泌体途径介导BVDV复制的研究[J].农业生物技术学报,2024,32(11):2662-2669,8.基金项目
自治区百名博士引进计划 （）
国家自然科学基金(31902271 （）
32060042) （）
自治区国际合作项目(2020E01006) （2020E01006）
自治区天山创新团队项目(2020D14005) （2020D14005）
新疆动物疫病研究重点实验室开放课题(2023KLA005) （2023KLA005）
