8TH1

Crystal Structure of the G3BP1 NTF2-like domain bound to the IDR1 of SARS-CoV-2 nucleocapsid protein D3L mutant

Summary for 8TH1

• Entry DOI: 10.2210/pdb8th1/pdb
• Descriptor: Ras GTPase-activating protein-binding protein 1, Nucleoprotein (3 entities in total)
• Functional Keywords: ntf2l covid, peptide binding protein, hydrolase-viral protein complex, hydrolase/viral protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 8
• Total formula weight: 84482.87

Authors

Hughes, M.P.,Taylor, J.P.,Yang, Z. (deposition date: 2023-07-13, release date: 2023-12-27, Last modification date: 2024-05-08)

Primary citation

Yang, Z.,Johnson, B.A.,Meliopoulos, V.A.,Ju, X.,Zhang, P.,Hughes, M.P.,Wu, J.,Koreski, K.P.,Clary, J.E.,Chang, T.C.,Wu, G.,Hixon, J.,Duffner, J.,Wong, K.,Lemieux, R.,Lokugamage, K.G.,Alvarado, R.E.,Crocquet-Valdes, P.A.,Walker, D.H.,Plante, K.S.,Plante, J.A.,Weaver, S.C.,Kim, H.J.,Meyers, R.,Schultz-Cherry, S.,Ding, Q.,Menachery, V.D.,Taylor, J.P.
Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity.
Cell Rep, 43:113965-113965, 2024

PubMed Abstract: G3BP1/2 are paralogous proteins that promote stress granule formation in response to cellular stresses, including viral infection. The nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) inhibits stress granule assembly and interacts with G3BP1/2 via an ITFG motif, including residue F17, in the N protein. Prior studies examining the impact of the G3PB1-N interaction on SARS-CoV-2 replication have produced inconsistent findings, and the role of this interaction in pathogenesis is unknown. Here, we use structural and biochemical analyses to define the residues required for G3BP1-N interaction and structure-guided mutagenesis to selectively disrupt this interaction. We find that N-F17A mutation causes highly specific loss of interaction with G3BP1/2. SARS-CoV-2 N-F17A fails to inhibit stress granule assembly in cells, has decreased viral replication, and causes decreased pathology in vivo. Further mechanistic studies indicate that the N-F17-mediated G3BP1-N interaction promotes infection by limiting sequestration of viral genomic RNA (gRNA) into stress granules.

PubMed: 38492217
DOI: 10.1016/j.celrep.2024.113965

Experimental method

X-RAY DIFFRACTION (1.8 Å)