1. SARS-CoV-2 Nucleocapsid Protein Targets a Conserved Surface Groove of the NTF2-like Domain of G3BP1
- Author
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Biswal, Mahamaya, Lu, Jiuwei, and Song, Jikui
- Subjects
Infectious Diseases ,Emerging Infectious Diseases ,Pneumonia ,Vaccine Related ,Lung ,Prevention ,Biodefense ,2.1 Biological and endogenous factors ,2.2 Factors relating to the physical environment ,Underpinning research ,Aetiology ,1.1 Normal biological development and functioning ,Infection ,Good Health and Well Being ,Coronavirus Nucleocapsid Proteins ,Crystallography ,DNA Helicases ,Humans ,Mutation ,Phosphoproteins ,Poly-ADP-Ribose Binding Proteins ,Protein Interaction Domains and Motifs ,RNA Helicases ,RNA Recognition Motif Proteins ,SARS-CoV-2 ,ras GTPase-activating protein-binding protein 1 ,SARS-CoV-2 nucleocapsid protein ,stress granule ,NTF2-like domain ,pathogen-host interaction ,Medicinal and Biomolecular Chemistry ,Biochemistry and Cell Biology ,Microbiology ,Biochemistry & Molecular Biology - Abstract
Stress granule (SG) formation mediated by Ras GTPase-activating protein-binding protein 1 (G3BP1) constitutes a key obstacle for viral replication, which makes G3BP1 a frequent target for viruses. For instance, the SARS-CoV-2 nucleocapsid (N) protein interacts with G3BP1 directly to suppress SG assembly and promote viral production. However, the molecular basis for the SARS-CoV-2 N - G3BP1 interaction remains elusive. Here we report biochemical and structural analyses of the SARS-CoV-2 N - G3BP1 interaction, revealing differential contributions of various regions of SARS-CoV-2 N to G3BP1 binding. The crystal structure of the NTF2-like domain of G3BP1 (G3BP1NTF2) in complex with a peptide derived from SARS-CoV-2 N (residues 1-25, N1-25) reveals that SARS-CoV-2 N1-25 occupies a conserved surface groove of G3BP1NTF2 via surface complementarity. We show that a φ-x-F (φ, hydrophobic residue) motif constitutes the primary determinant for G3BP1NTF2-targeting proteins, while the flanking sequence underpins diverse secondary interactions. We demonstrate that mutation of key interaction residues of the SARS-CoV-2 N1-25 - G3BP1NTF2 complex leads to disruption of the SARS-CoV-2 N - G3BP1 interaction in vitro. Together, these results provide a molecular basis of the strain-specific interaction between SARS-CoV-2 N and G3BP1, which has important implications for the development of novel therapeutic strategies against SARS-CoV-2 infection.
- Published
- 2022