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Despite the extensive attention MPXV has garnered, little is known about its clinical manifestations in humans. In this study, a high-throughput RNA sequencing (RNA-seq) and liquid chromatography-tandem mass spectrometry (LC-MS\/MS) approach was employed to investigate the transcriptional and metabolic responses of HEK293T cells to the MPXV A5L protein. RNA-seq analysis identified a total of 1473 differentially expressed genes (DEGs), comprising 911 upregulated and 562 downregulated genes. Additionally, LC-MS\/MS analysis revealed 185 cellular proteins with significantly altered abundance ratios that interact with the A5L protein. Here, we perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the transcriptome and proteome signatures of MPXV A5L-expressing HEK293T cells to gain insights into the virus proteins-host interplay. Transcriptomic analysis revealed that transfection of the MPXV A5L protein modulated genes primarily associated with the cell cycle, ribosome, and DNA replication. Proteomic analysis indicated that this protein predominantly interacted with host ribosomal proteins and cytoskeletal proteins. The combination of transcriptomic and proteomic analysis offers new perspectives for understanding the interaction between pathogens and hosts. Our research emphasizes the significant role of MPXV A5L in facilitating viral internalization and assembly, as well as its impact on the host\u2019s translation system.<\/jats:p>","DOI":"10.3390\/ijms26010398","type":"journal-article","created":{"date-parts":[[2025,1,6]],"date-time":"2025-01-06T08:08:52Z","timestamp":1736150932000},"page":"398","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Transcriptomic and Proteomic Analysis of Monkeypox Virus A5L-Expressing HEK293T Cells"],"prefix":"10.3390","volume":"26","author":[{"given":"Mingzhi","family":"Li","sequence":"first","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaqi","family":"Xiong","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Zhou","sequence":"additional","affiliation":[{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenyi","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengle","family":"Jia","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yihao","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nannan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanying","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Zhong","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhicheng","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruiying","family":"Li","sequence":"additional","affiliation":[{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunli","family":"Guo","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8128-1697","authenticated-orcid":false,"given":"Qi","family":"Peng","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingbao","family":"Kong","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330000, China"},{"name":"Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330000, China"},{"name":"College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1016\/S1473-3099(22)00228-6","article-title":"Clinical features and management of human monkeypox: A retrospective observational study in the UK","volume":"22","author":"Adler","year":"2022","journal-title":"Lancet Infect. 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