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Role of nucleocapsid protein of hantaviruses in intracellular traffic of viral glycoproteins

机译:汉坦病毒核衣壳蛋白在病毒糖蛋白胞内运输中的作用

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摘要

To understand the role of nucleocapsid protein (NP) of hantaviruses in viral assembly, the effect of NP on intracellular traffic of viral glycoproteins Gn and Gc was investigated. Double staining of viral and host proteins in Hantaan virus (HTNV)-infected Vero E6 cells showed that Gn and Gc were localized to cis-Golgi, in which virus particles are thought to be formed. When HTNV Gn and Gc were expressed by a plasmid encoding glycoprotein precursor (GPC), which is posttranslationaly cleaved into Gn and Gc, Gn was localized to cis-Golgi, whereas Gc showed diffuse distribution in the cytoplasm in 32.9% of Gc-positive cells. The ratio of the diffused Gc-positive cells was significantly decreased to 15.0% by co-expression of HTNV NP. Co-expression of HTNV GPC with NPs of other hantaviruses, such as Seoul virus, Puumala virus and Sin Nombre virus, also reduced the ratios of diffused Gc-positive cells to 13.5%, 25.2%, and 11.6%, respectively. Among amino- and carboxyl-terminally truncated HTNV NPs, NP75-429, NP116-429, NP1-333, NP1-233, and NP1-155 possessed activity to reduce the ratio of diffused Gc-positive cells, while NP155-429 and NP1-116 did not. NP30-429 has partial activity. These results indicate that amino acid region 116-155 of NP is important for the activity, although amino acid region 1-30 is partially related. Truncation of the HTNV Gc cytoplasmic tail caused an increase in diffused Gc-positive cells. In addition, the effect of coexpression of HTNV NP was weakened. These results suggest that HTNV NP has a role to promote Golgi localization of Gc through a mechanism possibly mediated by the Gc cytoplasmic tail.
机译:为了了解汉坦病毒的核衣壳蛋白(NP)在病毒装配中的作用,研究了NP对病毒糖蛋白Gn和Gc的细胞内运输的影响。汉坦病毒(HTNV)感染的Vero E6细胞中病毒和宿主蛋白的双重染色显示Gn和Gc定位于顺式高尔基体,其中病毒颗粒被认为形成了。当通过编码糖蛋白前体(GPC)的质粒表达HTNV Gn和Gc时,该质粒被翻译后裂解为Gn和Gc,Gn定位于顺式高尔基体,而Gc在32.9%的Gc阳性细胞中显示出分散在细胞质中。通过HTNV NP的共表达,弥散的Gc阳性细胞的比例显着降低至15.0%。 HTNV GPC与其他汉坦病毒(如汉城病毒,Puumala病毒和Sin Nombre病毒)的NPs共表达,还将弥散的Gc阳性细胞比例分别降低到13.5%,25.2%和11.6%。在氨基末端和羧基末端截短的HTNV NP中,NP75-429,NP116-429,NP1-333,NP1-233和NP1-155具有降低扩散的Gc阳性细胞比例的活性,而NP155-429和NP1 -116没有。 NP30-429具有部分活性。这些结果表明,尽管氨基酸区域1-30部分相关,但是NP的氨基酸区域116-155对于活性是重要的。 HTNV Gc细胞质尾部的截断导致弥散的Gc阳性细胞增加。另外,HTNV NP的共表达的作用减弱。这些结果表明,HTNV NP具有通过可能由Gc胞质尾部介导的机制来促进Gc的高尔基体定位的作用。

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