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 Plante; Scott C. Weaver; Hong Joo Kim; Rachel Meyers; Stacey Schultz-Cherry; Qiang Ding; Vineet D. Menachery; J. Paul Taylor

doi:10.1016/j.celrep.2024.113965

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. PlanteJessica Plante, Scott C. Weaver, Hong Joo Kim, Rachel Meyers, Stacey Schultz-Cherry, Qiang Ding, Vineet D. Menachery, J. Paul Taylor

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

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.

Original language	English (US)
Article number	113965
Pages (from-to)	113965
Journal	Cell Reports
Volume	43
Issue number	3
DOIs	https://doi.org/10.1016/j.celrep.2024.113965
State	Published - Mar 26 2024

Keywords

CP: Microbiology
G3BP
NTF2L domain
SARS-CoV-2 replication and pathogenesis
host-pathogen interaction
nucleocapsid protein
stress granule
vRNA sequestration

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/j.celrep.2024.113965

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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., ... Taylor, J. P. (2024). Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity. Cell Reports, 43(3), 113965. Article 113965. https://doi.org/10.1016/j.celrep.2024.113965

Yang, Z, Johnson, BA, Meliopoulos, VA, Ju, X, Zhang, P, Hughes, MP, Wu, J, Koreski, KP, Clary, JE, Chang, TC, Wu, G, Hixon, J, Duffner, J, Wong, K, Lemieux, R, Lokugamage, KG, Alvarado, RE, Crocquet-Valdes, PA, Walker, DH , Plante, KS, Plante, J, Weaver, SC, Kim, HJ, Meyers, R, Schultz-Cherry, S, Ding, Q, Menachery, VD & Taylor, JP 2024, 'Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity', Cell Reports, vol. 43, no. 3, 113965, pp. 113965. https://doi.org/10.1016/j.celrep.2024.113965

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title = "Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity",

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.",

keywords = "CP: Microbiology, G3BP, NTF2L domain, SARS-CoV-2 replication and pathogenesis, host-pathogen interaction, nucleocapsid protein, stress granule, vRNA sequestration",

author = "Zemin Yang and Johnson, {Bryan A.} and Meliopoulos, {Victoria A.} and Xiaohui Ju and Peipei Zhang and Hughes, {Michael P.} and Jinjun Wu and Koreski, {Kaitlin P.} and Clary, {Jemma E.} and Chang, {Ti Cheng} and Gang Wu and Jeff Hixon and Jay Duffner and Kathy Wong and Rene Lemieux and Lokugamage, {Kumari G.} and Alvarado, {R. Elias} and Crocquet-Valdes, {Patricia A.} and Walker, {David H.} and Plante, {Kenneth S.} and Jessica Plante and Weaver, {Scott C.} and Kim, {Hong Joo} and Rachel Meyers and Stacey Schultz-Cherry and Qiang Ding and Menachery, {Vineet D.} and Taylor, {J. Paul}",

note = "Publisher Copyright: {\textcopyright} 2024 The Author(s)",

year = "2024",

month = mar,

day = "26",

doi = "10.1016/j.celrep.2024.113965",

language = "English (US)",

volume = "43",

pages = "113965",

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TY - JOUR

T1 - Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity

AU - Yang, Zemin

AU - Johnson, Bryan A.

AU - Meliopoulos, Victoria A.

AU - Ju, Xiaohui

AU - Zhang, Peipei

AU - Hughes, Michael P.

AU - Wu, Jinjun

AU - Koreski, Kaitlin P.

AU - Clary, Jemma E.

AU - Chang, Ti Cheng

AU - Wu, Gang

AU - Hixon, Jeff

AU - Duffner, Jay

AU - Wong, Kathy

AU - Lemieux, Rene

AU - Lokugamage, Kumari G.

AU - Alvarado, R. Elias

AU - Crocquet-Valdes, Patricia A.

AU - Walker, David H.

AU - Plante, Kenneth S.

AU - Plante, Jessica

AU - Weaver, Scott C.

AU - Kim, Hong Joo

AU - Meyers, Rachel

AU - Schultz-Cherry, Stacey

AU - Ding, Qiang

AU - Menachery, Vineet D.

AU - Taylor, J. Paul

PY - 2024/3/26

Y1 - 2024/3/26

N2 - 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.

AB - 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.

KW - CP: Microbiology

KW - G3BP

KW - NTF2L domain

KW - SARS-CoV-2 replication and pathogenesis

KW - host-pathogen interaction

KW - nucleocapsid protein

KW - stress granule

KW - vRNA sequestration

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U2 - 10.1016/j.celrep.2024.113965

DO - 10.1016/j.celrep.2024.113965

M3 - Article

C2 - 38492217

AN - SCOPUS:85187986978

SN - 2211-1247

VL - 43

SP - 113965

JO - Cell Reports

JF - Cell Reports

IS - 3

M1 - 113965

ER -

Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity

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