Mouse Norovirus Infection Arrests Host Cell Translation Uncoupled from the Stress Granule-PKR-eIF2α Axis

• Published in: mBio Vol. 10; no. 3
• Main Authors: Fritzlar, Svenja, Aktepe, Turgut E, Chao, Yi-Wei, Kenney, Nathan D, McAllaster, Michael R, Wilen, Craig B, White, Peter A, Mackenzie, Jason M
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 18.06.2019
• Subjects: Animals; Caliciviridae Infections - immunology; Cytoplasmic Granules - immunology; Cytoplasmic Granules - virology; DNA Helicases - immunology; Editor's Pick; eIF-2 Kinase - immunology; eIF2α; Eukaryotic Initiation Factor-2 - immunology; Host-Microbe Biology; Host-Pathogen Interactions; Immune Evasion; Immunity, Innate; integrated stress response; Mice; mouse norovirus; Phosphorylation; Poly-ADP-Ribose Binding Proteins - immunology; Protein Biosynthesis; protein kinase R; protein translation; RNA Helicases - immunology; RNA Recognition Motif Proteins - immunology; stress granules; Viral Proteins - genetics; Viral Proteins - metabolism; Virus Replication; eIF2α; stress granules; protein translation; mouse norovirus; integrated stress response; protein kinase R
• ISSN: 2161-2129; 2150-7511
• DOI: 10.1128/mBio.00960-19

The integrated stress response (ISR) is a cellular response system activated upon different types of stresses, including viral infection, to restore cellular homeostasis. However, many viruses manipulate this response for their own advantage. In this study, we investigated the association between murine norovirus (MNV) infection and the ISR and demonstrate that MNV regulates the ISR by activating and recruiting key ISR host factors. We observed that during MNV infection, there is a progressive increase in phosphorylated eukaryotic initiation factor 2α (p-eIF2α), resulting in the suppression of host translation, and yet MNV translation still progresses under these conditions. Interestingly, the shutoff of host translation also impacts the translation of key signaling cytokines such as beta interferon, interleukin-6, and tumor necrosis factor alpha. Our subsequent analyses revealed that the phosphorylation of eIF2α was mediated via protein kinase R (PKR), but further investigation revealed that PKR activation, phosphorylation of eIF2α, and translational arrest were uncoupled during infection. We further observed that stress granules (SGs) are not induced during MNV infection and that MNV can restrict SG nucleation and formation. We observed that MNV recruited the key SG nucleating protein G3BP1 to its replication sites and intriguingly the silencing of G3BP1 negatively impacts MNV replication. Thus, it appears that MNV utilizes G3BP1 to enhance replication but equally to prevent SG formation, suggesting an anti-MNV property of SGs. Overall, this study highlights MNV manipulation of SGs, PKR, and translational control to regulate cytokine translation and to promote viral replication. Viruses hijack host machinery and regulate cellular homeostasis to actively replicate their genome, propagate, and cause disease. In retaliation, cells possess various defense mechanisms to detect, destroy, and clear infecting viruses, as well as signal to neighboring cells to inform them of the imminent threat. In this study, we demonstrate that the murine norovirus (MNV) infection stalls host protein translation and the production of antiviral and proinflammatory cytokines. However, virus replication and protein translation still ensue. We show that MNV further prevents the formation of cytoplasmic RNA granules, called stress granules (SGs), by recruiting the key host protein G3BP1 to the MNV replication complex, a recruitment that is crucial to establishing and maintaining virus replication. Thus, MNV promotes immune evasion of the virus by altering protein translation. Together, this evasion strategy delays innate immune responses to MNV infection and accelerates disease onset.