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Although MPXV attracts broad attention, there is a lack of adequate studies to elucidate pathogenic mechanisms associated with viral infections. In this study, a high-throughput RNA sequencing (RNA-seq) and liquid chromatography\u2013tandem mass spectrometry (LC-MS\/MS) approach was used to explore the transcriptional and metabolic responses of MPXV A23 protein to HEK293T cells. The protein\u2013protein interactions and signaling pathways were conducted by GO and KEGG analyses. The localization of A23 protein in HEK293T cells was detected by immunofluorescence. A total of 648 differentially expressed genes (DEGs) were identified in cells by RNA-Seq, including 314 upregulated genes and 334 downregulated genes. Additionally, liquid chromatography\u2013tandem mass spectrometry (LC-MS\/MS) detected 115 cellular proteins that interact with the A23 proteins. Transcriptomic sequencing analysis revealed that transfection of MPXV A23 protein modulated genes primarily associated with cellular apoptosis and DNA damage repair. Proteomic analysis indicated that this protein primarily interacted with host ribosomal proteins and histones. Following the identification of the nuclear localization sequence RKKR within the A23 protein, a truncated mutant A23\u0394RKKR was constructed to investigate the subcellular localization of A23 protein. The wild-type A23 protein exhibits a significantly higher nuclear-to-cytoplasmic ratio, exceeding 1.5, in contrast to the mutant A23\u0394RKKR, which has a ratio of approximately 1. Immunofluorescence assays showed that the A23 protein was mainly localized in the nucleus. The integration of transcriptomics and proteomics analysis provides a comprehensive understanding of the interaction between MPXV A23 protein and the host. Our findings highlight the potential role of this enzyme in suppressing host antiviral immune responses and modulating host gene expression.<\/jats:p>","DOI":"10.3390\/ijms25168678","type":"journal-article","created":{"date-parts":[[2024,8,9]],"date-time":"2024-08-09T07:59:07Z","timestamp":1723190347000},"page":"8678","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Combined Transcriptomic and Proteomic Analysis of Monkeypox Virus A23 Protein on HEK293T Cells"],"prefix":"10.3390","volume":"25","author":[{"given":"Yihao","family":"Wang","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":"Yihan","family":"Li","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":"Mingzhi","family":"Li","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":"Keyi","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":"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":"Ting","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"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9251-344X","authenticated-orcid":false,"given":"Yu","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":"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"}]},{"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"}]},{"given":"Meifeng","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 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