TMPRSS2 plays a crucial role in facilitating the entry of both the influenza virus and the SARSCoV-2 coronavirus into host cells.Recent studies have identified a guanine-rich sequence in the proximal promoter region o...TMPRSS2 plays a crucial role in facilitating the entry of both the influenza virus and the SARSCoV-2 coronavirus into host cells.Recent studies have identified a guanine-rich sequence in the proximal promoter region of the TMPRSS2 gene,which can form G-quadruplex structures(TMPRSS2-G4s)that are potential targets for small molecules to inhibit TMPRSS2 expression.However,the structural details of the major TMPRSS2-G4 and its complex with small molecules remain unknown,hindering the development of antiviral drugs targeting TMPRSS2-Gquadruplexes(G4s).This study reports the first high-resolution nuclear magnetic resonance(NMR)solution structure of the major TMPRSS2-G4,which consists of a three-tetrad core parallel-stranded G4.Both 3′and 5′flanking regions form well-defined capping structures stabilized by multiple hydrogen bonds.Importantly,we found that berberine,an antiviral alkaloid,strongly binds to the major TMPRSS2-G4 and determined its binding complex structure with TMPRSS2-G4 at a 2∶1 binding stoichiometry.Each berberine molecule recruits an adjacent flanking residue,forming a coplanar structure superimposed on two outer G-tetrads.Moreover,we demonstrated that the major TMPRSS2-G4 can stably form within a longer deoxyribonucleic acid(DNA)context and be targeted by small molecules to inhibit DNA polymerase activity.Overall,this study provides structural insights into the recognition mechanism of small molecules by the major TMPRSS2-G4 and may facilitate the development of novel antiviral therapeutics targeting TMPRSS2-G4.展开更多
基金supported by the National Natural Science Foundation of China(Nos.82322065,82173707,82204241)the Innovation and Entrepreneurship(Shuangchuang)Program of Jiang-su Province(2024)+3 种基金the Natural Science Foundation of Jiangsu Province(No.BK20221039)the Project Program of State Key Laboratory of Natural Medicines(China Pharmaceutical University,No.SKLNMZZ2024JS12)the Fundamental Research Funds for the Central Universities(No.2632025ZD06)the Scientific Research Foundation for High-level Faculty,China Pharmaceutical University(No.3150020065)。
文摘TMPRSS2 plays a crucial role in facilitating the entry of both the influenza virus and the SARSCoV-2 coronavirus into host cells.Recent studies have identified a guanine-rich sequence in the proximal promoter region of the TMPRSS2 gene,which can form G-quadruplex structures(TMPRSS2-G4s)that are potential targets for small molecules to inhibit TMPRSS2 expression.However,the structural details of the major TMPRSS2-G4 and its complex with small molecules remain unknown,hindering the development of antiviral drugs targeting TMPRSS2-Gquadruplexes(G4s).This study reports the first high-resolution nuclear magnetic resonance(NMR)solution structure of the major TMPRSS2-G4,which consists of a three-tetrad core parallel-stranded G4.Both 3′and 5′flanking regions form well-defined capping structures stabilized by multiple hydrogen bonds.Importantly,we found that berberine,an antiviral alkaloid,strongly binds to the major TMPRSS2-G4 and determined its binding complex structure with TMPRSS2-G4 at a 2∶1 binding stoichiometry.Each berberine molecule recruits an adjacent flanking residue,forming a coplanar structure superimposed on two outer G-tetrads.Moreover,we demonstrated that the major TMPRSS2-G4 can stably form within a longer deoxyribonucleic acid(DNA)context and be targeted by small molecules to inhibit DNA polymerase activity.Overall,this study provides structural insights into the recognition mechanism of small molecules by the major TMPRSS2-G4 and may facilitate the development of novel antiviral therapeutics targeting TMPRSS2-G4.
