Cell Reports (Mar 2024)
Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity
- Zemin Yang,
- Bryan A. Johnson,
- Victoria A. Meliopoulos,
- Xiaohui Ju,
- Peipei Zhang,
- Michael P. Hughes,
- Jinjun Wu,
- Kaitlin P. Koreski,
- Jemma E. Clary,
- Ti-Cheng Chang,
- Gang Wu,
- Jeff Hixon,
- Jay Duffner,
- Kathy Wong,
- Rene Lemieux,
- Kumari G. Lokugamage,
- R. Elias Alvarado,
- Patricia A. Crocquet-Valdes,
- David H. Walker,
- Kenneth S. Plante,
- Jessica A. Plante,
- Scott C. Weaver,
- Hong Joo Kim,
- Rachel Meyers,
- Stacey Schultz-Cherry,
- Qiang Ding,
- Vineet D. Menachery,
- J. Paul Taylor
Affiliations
- Zemin Yang
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA; Integrated Biomedical Sciences Program, University of Tennessee Health Science Center, Memphis, TN, USA
- Bryan A. Johnson
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Center for Tropical Diseases, University of Texas Medical Branch, Galveston, TX, USA
- Victoria A. Meliopoulos
- Department of Infectious Diseases, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Xiaohui Ju
- School of Medicine, Tsinghua University, Beijing, China
- Peipei Zhang
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Michael P. Hughes
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Jinjun Wu
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA; Integrated Biomedical Sciences Program, University of Tennessee Health Science Center, Memphis, TN, USA
- Kaitlin P. Koreski
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Jemma E. Clary
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Ti-Cheng Chang
- Center for Applied Bioinformatics, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Gang Wu
- Center for Applied Bioinformatics, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Jeff Hixon
- Faze Medicines, Cambridge, MA, USA
- Jay Duffner
- Faze Medicines, Cambridge, MA, USA
- Kathy Wong
- Faze Medicines, Cambridge, MA, USA
- Rene Lemieux
- Faze Medicines, Cambridge, MA, USA
- Kumari G. Lokugamage
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA
- R. Elias Alvarado
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA
- Patricia A. Crocquet-Valdes
- Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA
- David H. Walker
- Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA
- Kenneth S. Plante
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA
- Jessica A. Plante
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA
- Scott C. Weaver
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA
- Hong Joo Kim
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Rachel Meyers
- Faze Medicines, Cambridge, MA, USA
- Stacey Schultz-Cherry
- Department of Infectious Diseases, St. Jude Children’s Research Hospital, Memphis, TN, USA
- Qiang Ding
- School of Medicine, Tsinghua University, Beijing, China
- Vineet D. Menachery
- Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA; Corresponding author
- J. Paul Taylor
- Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, Memphis, TN, USA; Howard Hughes Medical Institute, Chevy Chase, MD, USA; Corresponding author
- Journal volume & issue
-
Vol. 43,
no. 3
p. 113965
Abstract
Summary: 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.
