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ATP 合酶亚基 ATP5B 与 TGEV Nsp2 相互作用，充当 TGEV 复制的负调节剂。

ATP synthase subunit ATP5B interacts with TGEV Nsp2 and acts as a negative regulator of TGEV replication.

机构信息

College of Veterinary Medicine, Northeast Agricultural University, Harbin, China.

China Ministry of Agriculture Key Laboratory of Animal Pathogen Biology, Northeastern Science Inspection Station, Harbin, China.

出版信息

Virulence. 2024 Dec;15(1):2397492. doi: 10.1080/21505594.2024.2397492. Epub 2024 Sep 9.

DOI:10.1080/21505594.2024.2397492
PMID:39239724
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11385163/
Abstract

Coronavirus nonstructural protein 2 (Nsp2) is regarded as a virulence determinant and plays a critical role in virus replication, and innate immunity. Screening and identifying host cell proteins that interact with viral proteins is an effective way to reveal the functions of viral proteins. In this study, the host proteins that interacted with transmissible gastroenteritis virus (TGEV) Nsp2 were identified using immunoprecipitation combined with LC-MS/MS. 77 host cell proteins were identified as putative Nsp2 interaction host cell proteins and a protein-protein interaction (PPI) was constructed. The identified proteins were found to be associated with various subcellular locations and functional categories through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. It is hypothesized that the host cell proteins interacting with TGEV Nsp2 are mainly involved in the formation of the cytoplasmic translation initiation complex, mRNA binding, ribosomes, and proteasomes. Among these, the ATP5B, a core subunit of the mitochondrial ATP synthase was further studied. The Coimmunoprecipitation (Co-IP) and indirect immunofluorescence (IFA) results confirmed that TGEV Nsp2 interacted with ATP5B. Furthermore, the downregulation of ATP5B expression was found to promote TGEV replication, suggesting that ATP5B might function as a negative regulator of TGEV replication. Collectively, our results offer additional insights into the functions of Nsp2 and provide a novel antiviral target against TGEV.

摘要

冠状病毒非结构蛋白 2（Nsp2）被认为是一种毒力决定因素，在病毒复制和固有免疫中发挥关键作用。筛选和鉴定与病毒蛋白相互作用的宿主细胞蛋白是揭示病毒蛋白功能的有效方法。在这项研究中，使用免疫沉淀结合 LC-MS/MS 鉴定了与传染性胃肠炎病毒（TGEV）Nsp2 相互作用的宿主蛋白。鉴定了 77 种宿主细胞蛋白作为推定的 Nsp2 相互作用宿主细胞蛋白，并构建了蛋白质-蛋白质相互作用（PPI）网络。通过基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析发现，鉴定的蛋白与各种亚细胞位置和功能类别相关。假设与 TGEV Nsp2 相互作用的宿主细胞蛋白主要参与细胞质翻译起始复合物、mRNA 结合、核糖体和蛋白酶体的形成。其中，ATP5B（线粒体 ATP 合酶的核心亚基）进一步进行了研究。共免疫沉淀（Co-IP）和间接免疫荧光（IFA）结果证实 TGEV Nsp2 与 ATP5B 相互作用。此外，下调 ATP5B 的表达被发现可促进 TGEV 的复制，表明 ATP5B 可能作为 TGEV 复制的负调控因子发挥作用。总之，我们的研究结果为 Nsp2 的功能提供了更多的见解，并为针对 TGEV 的新型抗病毒靶点提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/6a815dcfba4c/KVIR_A_2397492_F0008_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/e06126f6703f/KVIR_A_2397492_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/5f36b5f5ad81/KVIR_A_2397492_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/b5ee410286ea/KVIR_A_2397492_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/3431a55fd8a0/KVIR_A_2397492_F0004_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/cb3545e8f4a2/KVIR_A_2397492_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/45ee76c7d018/KVIR_A_2397492_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/499ef97c92b6/KVIR_A_2397492_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/6a815dcfba4c/KVIR_A_2397492_F0008_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/e06126f6703f/KVIR_A_2397492_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/5f36b5f5ad81/KVIR_A_2397492_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/b5ee410286ea/KVIR_A_2397492_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/3431a55fd8a0/KVIR_A_2397492_F0004_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/cb3545e8f4a2/KVIR_A_2397492_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/45ee76c7d018/KVIR_A_2397492_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/499ef97c92b6/KVIR_A_2397492_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa41/11385163/6a815dcfba4c/KVIR_A_2397492_F0008_OC.jpg

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