Retinoic Acid Inducible Gene I and Protein Kinase R, but Not Stress Granules, Mediate the Proinflammatory Response to Yellow Fever Virus
| Autor: | Maxime Chazal, Sarah Lesage, Takashi Fujita, Ségolène Gracias, Eliane F. Meurs, Laura Sinigaglia, Frédéric Tangy, Daniela Bruni, Felix Streicher, Guillaume Beauclair, Salomé Bourgeau, Nolwenn Jouvenet |
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| Přispěvatelé: | Département de Virologie - Department of Virology, Institut Pasteur [Paris] (IP), Centre National de la Recherche Scientifique (CNRS), Signalisation antivirale - Virus sensing and signaling, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Université Paris Cité (UPCité), Génomique virale et vaccination, Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Kyoto University, Hépacivirus et Immunité innée, ANR-16-CE15002501, Agence Nationale de la Recherche, ANR-16-CE15-0025,Viro-Storm,Mécanismes de production incontrôlée de cytokines au cours de l'infection virale(2016), Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Université de Paris (UP), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Kyoto University [Kyoto] |
| Rok vydání: | 2020 |
| Předmět: |
Small interfering RNA
MESH: Poly-ADP-Ribose Binding Proteins viruses MESH: DNA Helicases MESH: DEAD Box Protein 58 MESH: Hepatocytes eIF-2 Kinase flavivirus MESH: RNA Small Interfering Stress granule assembly MESH: Animals RNA Small Interfering Receptors Immunologic MESH: Carcinoma Hepatocellular Poly-ADP-Ribose Binding Proteins innate immunity MESH: Haplorhini 0303 health sciences MESH: Cytokines MESH: RNA Helicases 030302 biochemistry & molecular biology RNA-Binding Proteins Haplorhini liver inflammation 3. Good health Cell biology Virus-Cell Interactions interferons RNA Recognition Motif Proteins MESH: RNA Viral Gene Knockdown Techniques [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology MESH: T-Cell Intracellular Antigen-1 Cytokines DEAD Box Protein 58 RNA Viral Yellow fever virus RNA Helicases stress granules MESH: Cell Line Tumor Carcinoma Hepatocellular Immunology Biology Microbiology Virus Proinflammatory cytokine Cell Line MESH: eIF-2 Kinase 03 medical and health sciences Stress granule Virology Cell Line Tumor Animals Humans 030304 developmental biology MESH: Adaptor Proteins Signal Transducing Adaptor Proteins Signal Transducing MESH: Humans MESH: Transcriptome pattern recognition receptors DNA Helicases RNA virus biology.organism_classification Protein kinase R MESH: Gene Knockdown Techniques MESH: Yellow fever virus MESH: Cell Line T-Cell Intracellular Antigen-1 MESH: RNA-Binding Proteins Insect Science MESH: RNA Recognition Motif Proteins Hepatocytes Cytokine secretion Transcriptome |
| Zdroj: | J Virol Journal of Virology Journal of Virology, 2020, 94 (22), pp.e00403-20. ⟨10.1128/JVI.00403-20⟩ Journal of Virology, American Society for Microbiology, 2020, 94 (22), pp.e00403-20. ⟨10.1128/JVI.00403-20⟩ |
| ISSN: | 1098-5514 0022-538X |
| Popis: | International audience; Yellow fever virus (YFV) is an RNA virus primarily targeting the liver. Severe YF cases are responsible for hemorrhagic fever, plausibly precipitated by excessive proinflammatory cytokine response. Pathogen recognition receptors (PRRs), such as the cytoplasmic retinoic acid inducible gene I (RIG-I)-like receptors (RLRs), and the viral RNA sensor protein kinase R (PKR), are known to initiate a proinflammatory response upon recognition of viral genomes. Here, we sought to reveal the main determinants responsible for the acute cytokine expression occurring in human hepatocytes following YFV infection. Using a RIG-I-defective human hepatoma cell line, we found that RIG-I largely contributes to cytokine secretion upon YFV infection. In infected RIG-I-proficient hepatoma cells, RIG-I was localized in stress granules. These granules are large aggregates of stalled translation preinitiation complexes known to concentrate RLRs and PKR and are so far recognized as hubs orchestrating RNA virus sensing. Stable knockdown of PKR in hepatoma cells revealed that PKR contributes to both stress granule formation and cytokine induction upon YFV infection. However, stress granule disruption did not affect the cytokine response to YFV infection, as assessed by small interfering RNA (siRNA)-knockdown-mediated inhibition of stress granule assembly. Finally, no viral RNA was detected in stress granules using a fluorescence hybridization approach coupled with immunofluorescence. Our findings suggest that both RIG-I and PKR mediate proinflammatory cytokine induction in YFV-infected hepatocytes, in a stress granule-independent manner. Therefore, by showing the uncoupling of the cytokine response from the stress granule formation, our model challenges the current view in which stress granules are required for the mounting of the acute antiviral response. Yellow fever is a mosquito-borne acute hemorrhagic disease caused by yellow fever virus (YFV). The mechanisms responsible for its pathogenesis remain largely unknown, although increased inflammation has been linked to worsened outcome. YFV targets the liver, where it primarily infects hepatocytes. We found that two RNA-sensing proteins, RIG-I and PKR, participate in the induction of proinflammatory mediators in human hepatocytes infected with YFV. We show that YFV infection promotes the formation of cytoplasmic structures, termed stress granules, in a PKR- but not RIG-I-dependent manner. While stress granules were previously postulated to be essential platforms for immune activation, we found that they are not required for the production of proinflammatory mediators upon YFV infection. Collectively, our work uncovered molecular events triggered by the replication of YFV, which could prove instrumental in clarifying the pathogenesis of the disease, with possible repercussions for disease management. |
| Databáze: | OpenAIRE |
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