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 |
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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 |
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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) |
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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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