Date of Award

6-2023

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Program

Biomedical Sciences

Track

Neuroscience

Research Advisor

J. Paul Taylor, MD, PhD

Committee

William E. Armstrong, PhD; Mondira Kundu, MD, PhD; Peter J. McKinnon, PhD; Tanja Mittag, PhD

Keywords

G3BP, Nucleocapsid Protein, SARS-CoV-2, Stress Granule, Viral Genomic RNA

Abstract

G3BP1/2 are RNA-binding proteins that promote condensation to form stress granules in response to various cellular stresses, including viral infection. G3BP1/2 are prominent interactors of the nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the functional consequences of the G3BP-N interaction in the context of viral infection have remained unclear. Here we used structural and biochemical analyses to define the residues required for G3BP1-N interaction, followed by structure-guided mutagenesis of G3BP1 and N to selectively disrupt their interaction. We found that mutation of N-F17 led to selective loss of interaction with G3BP1 and consequent failure of the N protein to disrupt stress granule assembly. Introduction of SARS-CoV-2 bearing an F17A mutation in an in vivo hamster model resulted in significant decreases in viral replication, symptom severity, and pathology, suggesting that the G3BP1-N interaction promotes viral replication by suppressing the ability of G3BP1 to form stress granules.

Declaration of Authorship

Declaration of Authorship is included in the supplemental files.

ORCID

https://orcid.org/0009-0001-0590-2180

DOI

10.21007/etd.cghs.2023.0633

2023-020-Yang-DOA.pdf (108 kB)
Declaration of Authorship

Available for download on Sunday, July 06, 2025

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