The present study was conducted to determine whether avian reovirus (ARV) activates the phosphatidylinositol 3-kinase-dependent Akt (PI3K/Akt) pathway according to the PXXP or YXXXM motifs of σA and σNS proteins. Gene splicing by overlap extension PCR was used to change the PXXP or YXXXM motifs of the σA and σNS genes. Plasmid constructs that contain mutant σA and σNS genes were generated and transfected into Vero cells, and the expression levels of the corresponding genes were quantified according to immunofluorescence and Western blot analyses. The Akt phosphorylation (P-Akt) profile of the transfected Vero cells was examined by flow cytometry and Western blot. The results showed that the σA and σNS genes were expressed in the Vero cells, and P-Akt expression in the σA mutant groups (amino acids 110-114 and 114-117) was markedly decreased. The results indicated that the σA protein of ARV activates the PI3K/Akt pathway via the PXXP motif. The results of this study reveal the mechanisms by which ARV manipulates the cellular signal transduction pathways, which may provide new ideas for novel drug targets.

中文翻译：

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通过禽呼肠孤病毒的σA和σNS蛋白基序研究PI3K / Akt途径的激活。

进行本研究，以确定是否根据σA和σNS蛋白的PXXP或YXXXM图案，确定禽呼肠孤病毒（ARV）是否激活了磷脂酰肌醇3激酶依赖性Akt（PI3K / Akt）途径。通过重叠延伸PCR进行的基因剪接用于改变σA和σNS基因的PXXP或YXXXM基序。产生包含突变的σA和σNS基因的质粒构建体，并将其转染到Vero细胞中，并根据免疫荧光和Western印迹分析定量相应基因的表达水平。通过流式细胞术和Western印迹检查转染的Vero细胞的Akt磷酸化（P-Akt）概况。结果表明，在Vero细胞中表达了σA和σNS基因，并且在σA突变体组（氨基酸110-114和114-117）中的P-Akt表达显着降低。结果表明，ARV的σA蛋白通过PXXP基序激活PI3K / Akt途径。这项研究的结果揭示了抗逆转录病毒操纵细胞信号转导途径的机制，这可能为新型药物靶标提供新的思路。