狂犬病病毒M蛋白胞内合成和分布初步研究

    Preliminary Study on Intracellular Synthesis and Translocation of Rabies Virus M Protein

    • 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N2A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N2A或BHK细胞的TCID50,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N2A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。

       

      Abstract:
      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 N2A and BHK cells were studied to determine the intracellular synthesis pathway and translocation of RABV M proteins.
      Method TCID50 of different RABV strains (CVS-11, SRV-9 and PB4) in N2A 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.

       

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