ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/090/SRR31885690/SRR31885690_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/092/SRR31885692/SRR31885692_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/094/SRR31885694/SRR31885694_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/093/SRR31885693/SRR31885693_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/093/SRR31885693/SRR31885693_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/095/SRR31885695/SRR31885695_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/091/SRR31885691/SRR31885691_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/090/SRR31885690/SRR31885690_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/092/SRR31885692/SRR31885692_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/095/SRR31885695/SRR31885695_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/094/SRR31885694/SRR31885694_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR318/091/SRR31885691/SRR31885691_1.fastq.gzprimaryOK200GenomicsUniversity of Pittsburghhttps://www.ebi.ac.uk/ena/browser/view/PRJNA1206177NSP7 (Non-Structural Protein 7) of SARS-CoV-2 is a crucial component for viral replication and transcription. In this study, we reveal that the host E3 ubiquitin ligase FBXO5 suppresses viral replication by mediating NSP7 lysine-48-linked ubiquitination and subsequent proteasomal degradation. Interestingly, we have also determined that NSP7 expression impairs the host antiviral response by inhibiting the ISGylation of melanoma differentiation-associated protein 5 (MDA5), a key sensor for viral RNA. Through an unbiased esiRNA screen, we identified that NSP7 ubiquitination is co-regulated by beta-TrCP1 and the kinase TAF1. Additionally, we identified a small-molecule FBXO5 stabilizer that disrupts the beta-TrCP1-FBXO5 interaction, thereby markedly enhancing NSP7 degradation and effectively mitigating SARS-CoV-2 infection. Taken together, our findings reveal a novel mechanism for NSP7 regulation and suggest that small-molecule activators of the E3 ubiquitin ligase FBXO5 represent a promising new class of host-directed antiviral therapies.ENAfalseNSP7 Molecule Degrader Suppresses Coronavirus Infection2025-01-072025-01-07PRJNA1206177