UNASSIGNED: The gene expression profiles of pCAGGS-HA-F3L transfected HEK293T cells were analyzed using RNA-seq. Proteomics was used to identify and study proteins that interact with F3L. Real-time PCR was used to detect mRNA levels of several differentially expressed genes (DEGs) in HEK293T cells (or Vero cells) after the expression of F3 protein.
UNASSIGNED: A total of 14,822 genes were obtained in cells by RNA-Seq and 1,672 DEGs were identified, including 1,156 up-regulated genes and 516 down-regulated genes. A total of 27 cellular proteins interacting with F3 proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and 19 cellular proteins with large differences in abundance ratios were considered to be candidate cellular proteins. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that the DEGs were significantly enriched in immune-related pathways, including type I interferon signaling pathway, response to virus, RIG-I-like receptor signaling pathway, NOD-like receptor signaling pathway, etc. Moreover, some selected DEGs were further confirmed by real-time PCR and the results were consistent with the transcriptome data. Proteomics data show that cellular proteins interacting with F3 proteins are mainly related to RNA splicing and protein translation.
UNASSIGNED: Our analysis of transcriptomic and proteomic data showed that (1) F3L up-regulates the transcript levels of key genes in the innate immune signaling pathway, such as RIGI, MDA5, IRF5, IRF7, IRF9, ISG15, IFNA14, and elicits a broad spectrum of antiviral immune responses in the host. F3L also increases the expression of the FOS and JNK genes while decreasing the expression of TNFR2, these factors may ultimately induce apoptosis. (2) F3 protein interacts with host proteins involved in RNA splicing and protein translation, such as SNRNP70, POLR2H, HNRNPA1, DDX17, etc. The findings of this study shed light on the function of the F3 protein.
使用RNA-seq分析pCAGGS-HA-F3L转染的HEK293T细胞的基因表达谱。蛋白质组学用于鉴定和研究与F3L相互作用的蛋白质。采用实时PCR检测F3蛋白表达后HEK293T细胞(或Vero细胞)中几种差异表达基因(DEGs)的mRNA水平。
■通过RNA-Seq在细胞中总共获得了14,822个基因,并鉴定了1,672个DEGs,包括1,156个上调基因和516个下调基因。通过液相色谱-串联质谱法(LC-MS/MS)鉴定出与F3蛋白相互作用的27种细胞蛋白,19种丰度比差异较大的细胞蛋白被认为是候选细胞蛋白。基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析表明,DEGs在免疫相关途径中显著富集,包括I型干扰素信号通路,对病毒的反应,RIG-I样受体信号通路,NOD样受体信号通路,等。此外,一些选定的DEGs通过实时PCR进一步证实,结果与转录组数据一致.蛋白质组学数据表明,细胞蛋白与F3蛋白相互作用主要与RNA剪接和蛋白质翻译有关。
■我们对转录组和蛋白质组数据的分析表明,(1)F3L上调了先天免疫信号通路中关键基因的转录水平,比如RIGI,MDA5、IRF5、IRF7、IRF9、ISG15、IFNA14和在宿主中引发广谱的抗病毒免疫应答。F3L还增加了FOS和JNK基因的表达,同时降低了TNFR2的表达,这些因素可能最终诱导细胞凋亡。(2)F3蛋白与参与RNA剪接和蛋白质翻译的宿主蛋白相互作用,如SNRNP70,POLR2H,HNRNPA1、DDX17等.这项研究的发现揭示了F3蛋白的功能。