mBio. 2023 Feb 28
Ying Li, Quanwei Yu, Ridong Huang, Hai Chen, Hequan Ren, Lingling Ma,Yang He, and Weimin Li
Products used in the paper Details Operation
lentiviruses packaging For co-IP, RNA-seq, ChIPqPCR, and apoptosis assays, SUD2core was generated into a pcDNA3.1-3FLAG vector (Thermo Fisher Scientific catalog number V79020) and packed with lentiviruses by PackGene Co. Request Quote

Research Field: Covid-related

Keywords: DNA topology, epithelial cells, gene expression, protein-DNA interactions, protein-protein interactions

AAV Serotype: AAV2/8


The molecular mechanisms underlying how SUD2 recruits other proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to exert its G-quadruplex (G4)-dependent pathogenic function is unknown. Herein, Nsp5 was singled out as a binding partner of the SUD2-N+M domains (SUD2core) with high affinity, through the surface located crossing these two domains. Biochemical and fluorescent assays demonstrated that this complex also formed in the nucleus of living host cells. Moreover, the SUD2core-Nsp5 complex displayed significantly enhanced selective binding affinity for the G4 structure in the BclII promoter than did SUD2core alone. This increased stability exhibited by the tertiary complex was rationalized by AlphaFold2 and molecular dynamics analysis. In line with these molecular interactions, downregulation of BclII and subsequent augmented apoptosis of respiratory cells were both observed. These results provide novel information and a new avenue to explore therapeutic strategies targeting SARS-CoV-2.

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