鸡 dsRNA 结合蛋白 hnRNPU 的特性与功能分析。
Characterization and functional analysis of chicken dsRNA binding protein hnRNPU.
机构信息
Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Zhimin Street, Qingshan Lake, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Science and Technology, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.
Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Zhimin Street, Qingshan Lake, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Science and Technology, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.
出版信息
Dev Comp Immunol. 2023 Jan;138:104521. doi: 10.1016/j.dci.2022.104521. Epub 2022 Aug 28.
In mammals, heterogeneous ribonucleoprotein U (hnRNPU), also named as nuclear matrix protein-nuclear scaffold attachment factor (SAFA), was originally identified as a DNA/RNA interactor protein. It has been reported that human hnRNPU facilitates IFN-β generation after vesicular stomatitis virus (VSV) infection. Nevertheless, the role of chicken hnRNPU (chhnRNPU) in IFN-β regulation as well as in infectious bursal diseases virus (IBDV) replication is still unclear. Here, we found that chhnRNPU inhibits IFN-β production via interacting with MDA5 and MAVS, and facilitates IBDV replication via associating with genomic dsRNA of IBDV. Firstly, chicken hnRNPU (chhnRNPU) was widely expressed in different tissues of chickens and was distributed in the nucleus of DF-1 cells. Overexpression of chhnRNPU significantly suppresses IFN-β promoter activities induced by MDA5 and MAVS. Additionally, immunoprecipitated by dsRNA antibodies, which followed LC-MS analysis demonstrate that chhnRNPU is a partner of viral genomic dsRNA. chhnRNPU is translocated from nucleus to cytosol to co-localize with replication complex of IBDV after IBDV infection. Over-expression of chhnRNPU significantly promotes IBDV replication, which was determined by western blotting, qRT-PCR and TCID assay. Furthermore, knock down chhnRNPU by siRNA remarkably facilitates IFN-β production, and inhibits IBDV proliferation. These data collectively reveal that chhnRNPU positively regulates IBDV replication via negatively regulating IFN-β response.
在哺乳动物中，异质核糖核蛋白 U（hnRNPU），也称为核基质蛋白-核骨架附着因子（SAFA），最初被鉴定为 DNA/RNA 相互作用蛋白。据报道，人 hnRNPU 有助于水疱性口炎病毒（VSV）感染后 IFN-β 的产生。然而，鸡 hnRNPU（chhnRNPU）在 IFN-β 调节以及传染性法氏囊病病毒（IBDV）复制中的作用尚不清楚。在这里，我们发现 chhnRNPU 通过与 MDA5 和 MAVS 相互作用抑制 IFN-β 的产生，并通过与 IBDV 的基因组 dsRNA 结合促进 IBDV 的复制。首先，鸡 hnRNPU（chhnRNPU）在鸡的不同组织中广泛表达，并分布在 DF-1 细胞的核内。chhnRNPU 的过表达显著抑制 MDA5 和 MAVS 诱导的 IFN-β 启动子活性。此外，用 dsRNA 抗体进行免疫沉淀，随后进行 LC-MS 分析表明，chhnRNPU 是病毒基因组 dsRNA 的伴侣。IBDV 感染后，chhnRNPU 从核内易位到细胞质，并与 IBDV 的复制复合物共定位。chhnRNPU 的过表达显著促进 IBDV 的复制，这通过 Western blot、qRT-PCR 和 TCID 测定来确定。此外，通过 siRNA 敲低 chhnRNPU 显著促进 IFN-β 的产生，并抑制 IBDV 的增殖。这些数据共同表明，chhnRNPU 通过负调控 IFN-β 反应正向调节 IBDV 复制。
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