摘要
【目的】狂犬病病毒(Rabies virus,RABV)是一种主要由核蛋白(N)、磷蛋白(P)、基质蛋白(M)、糖蛋白(G)、依赖RNA的RNA聚合酶蛋白(L)5种结构蛋白组成的高度嗜神经性病毒。RABV基因组RNA与N蛋白、P蛋白和L蛋白组成核糖核蛋白复合体(RNP),M蛋白在调控RABV病毒结构蛋白的合成和装配过程中起重要作用。但RABV不同毒株M蛋白胞内合成、分布场所及其是否存在差异尚不清楚。研究不同RABV毒株M蛋白与内质网、高尔基体以及M蛋白与G蛋白、M蛋白与RNP在N_(2)A和BHK细胞内的共定位,以期确定RABV M蛋白的胞内合成途径和分布场所。【方法】测定RABV不同毒株(CVS-11、SRV-9、PB4)在N_(2)A或BHK细胞的TCID_(50),以相同的MOI接毒,通过免疫荧光研究M蛋白与内质网、高尔基体的共定位,以及M蛋白与G蛋白、M蛋白与RNP(为N蛋白代表)的共定位。【结果】RABV不同毒株效价存在一定差异。在N_(2)A和BHK细胞中,不同RABV毒株的M蛋白与内质网、高尔基体均存在共定位;M蛋白与G蛋白、M蛋白与N蛋白也存在共定位,但M蛋白与G蛋白主要共定位于内质网、高尔基体上,而M蛋白与N蛋白主要共定位在胞浆中。【结论】RABV M蛋白在胞内主要通过内质网-高尔基体途径合成加工。研究结果明确了RABV M蛋白胞内合成分布部位,为进一步研究RABV病毒粒子的装配和出芽过程丰富了数据。
【Objective】Rabies virus(RABV)is a highly neurotropic virus,which composes of five structural proteins,such as nucleoprotein(N),phosphoprotein(P),matrix protein(M),glycoprotein(G)and RNA-dependent RNA polymerase(L).The RABV genomic RNA,combining with N,P and L proteins,constitute the ribonucleoprotein complex(RNP).M proteins plays a particular role in regulating the synthesis and assembly of the structural proteins of RABV.However,it is not clear where the intracellular synthesis and translocation sites of M proteins are and whether there are differences in M proteins in different cell lines among RABV strains.The co-localizations of M protein with ER and Golgi,M protein with G protein and M protein with RNP in N_(2)A and BHK cells were studied to determine the intracellular synthesis pathway and translocation of RABV M proteins.【Method】TCID_(50) of different RABV strains(CVS-11,SRV-9 and PB4)in N_(2)A or BHK cells were determined.Co-localization of M protein with ER and Golgi as well as with G protein and RNP(represented by N protein)in ER and Golgi apparatuses were performed by Immunofluorescent.【Result】The titers in the different RABV strains had certain differences.All the M proteins in different RABV strains were co-localized with ER and Golgi.M protein also co-localized with G protein as well as with N protein;however,the co-localization of M protein and G protein was mainly in ER and Golgi,while that of M protein and N protein in cytoplasm.【Conclusion】The synthesis and processing of the RABV M proteins were carried out through ER-Golgi pathways.The study results confirmed the synthesis and translocation cellular sites of RABV M proteins,and enriched the data for assembly and budding process of RABV virions.
作者
赵维荣
赵铭昕
徐婧
张丹玮
郭艺迪
关振宏
段铭
张茂林
ZHAO Weirong;ZHAO Mingxin;XU Jing;ZHANG Danwei;GUO Yidi;GUAN Zhenhong;DUAN Ming;ZHANG Maolin(Institute of Zoonosis,Jilin University/Key Laboratory of Zoonosis,Ministry of Education,Changchun 130062,China;College of Veterinary Medicine,Jilin University,Changchun 130062,China)
出处
《广东农业科学》
CAS
2021年第4期111-118,共8页
Guangdong Agricultural Sciences
基金
国家重点研发计划项目(2016YFD0500402)
国家自然科学基金(31972712,31702238)。
关键词
狂犬病病毒
M蛋白
内质网
高尔基体
rabies virus
M protein
endoplasmic reticulum(ER)
Golgi
