文章摘要
赵维荣,赵铭昕,徐 婧,张丹玮,郭艺迪,关振宏,段 铭,张茂林.狂犬病病毒 M 蛋白胞内合成和分布初步研究[J].广东农业科学,2021,48(4):111-118
查看全文    HTML 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
  
DOI:10.16768/j.issn.1004-874X.2021.04.016
中文关键词: 狂犬病病毒  M 蛋白  内质网  高尔基体
英文关键词: rabies virus  M protein  endoplasmic reticulum (ER)  Golgi
基金项目:国家重点研发计划项目(2016YFD0500402);国家自然科学基金(31972712,31702238)
作者单位
赵维荣,赵铭昕,徐 婧,张丹玮,郭艺迪,关振宏,段 铭,张茂林 1. 吉林大学人兽共患病研究所 / 人兽共患病研究教育部重点实验室吉林 长春 130062 2. 吉林大学动物医学学院吉林 长春 130062 
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中文摘要:
      【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 N2 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 N2 A 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 N2 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。
英文摘要:
      【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 N2 A 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 N2 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.
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