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Innate Immune Priming by cGAS as a Preparatory Countermeasure Against RNA Virus Infection

Innate Immune Priming by cGAS as a Preparatory Countermeasure Against RNA Virus Infection

来源:bioRxiv_logobioRxiv
英文摘要

Abstract The detection of nucleic acids by pattern recognition receptors is an ancient and conserved component of the innate immune system. Notably, RNA virus genomes are sensed by mammalian cytosolic RIG-I–like receptors, thereby activating interferon-stimulated gene (ISG) expression to restrict viral replication. However, recent evidence indicates that the cGAS-STING DNA sensing pathway also protects against RNA viruses. So far, the mechanisms responsible for DNA sensing of RNA viruses, which replicate without known DNA intermediates, remain unclear. By using cGAS gene knockout and reconstitution in human and mouse cell cultures, we discovered that DNA sensing and cGAMP synthase activities are required for cGAS-mediated restriction of vesicular stomatitis virus and Sindbis virus. The level of cGAMP produced in response to RNA virus infection was below the threshold of detection, suggesting that only transient and/or low levels of cGAMP are produced during RNA virus infections. To clarify the DNA ligands that activate cGAS activity, we confirmed that cGAS binds mitochondrial DNA in the cytosol of both uninfected and infected cells; however, the amount of cGAS-associated mitochondrial DNA did not change in response to virus infection. Rather, a variety of pre-existing cytosolic DNAs, including mitochondrial DNA and endogenous cDNAs, may serve as stimuli for basal cGAS activation. Importantly, cGAS knockout and reconstitution experiments demonstrated that cGAS drives low-level ISG expression at steady state. We propose that cGAS-STING restricts RNA viruses by promoting a preparatory immune activation state within cells, likely primed by endogenous cellular DNA ligands. ImportanceMany medically important RNA viruses are restricted by the cGAS-STING DNA-sensing pathway of innate immune activation. Since these viruses replicate without DNA intermediates, it is unclear what DNA ligand(s) are responsible for triggering this pathway. We show here that cGAS’s DNA binding and signaling activities are required for RNA virus restriction, similar to the mechanisms by which it restricts DNA viruses. Furthermore, we confirmed that cGAS continuously binds host DNA, which was unaffected by RNA virus infection. Finally, cGAS expression correlated with the low-level expression of interferon-stimulated genes in uninfected cells, both in vitro and in vivo. We propose that cGAS-mediated sensing of endogenous DNA ligands contributes to RNA virus restriction by establishing a baseline of innate immune activation.

Parker Michael T.、Gopinath Smita、Perez Corey E.、Crawford Jason M.、Iwasaki Akiko、Lindenbach Brett D.、Linehan Melissa M.

Department of Microbial Pathogenesis, Yale University School of Medicine||Department of Immunobiology, Yale University School of MedicineDepartment of Immunobiology, Yale University School of MedicineDepartment of Chemistry, Yale University||Chemical Biology Institute, Yale UniversityDepartment of Microbial Pathogenesis, Yale University School of Medicine||Department of Chemistry, Yale University||Chemical Biology Institute, Yale UniversityDepartment of Immunobiology, Yale University School of Medicine||Howard Hughes Medical Institute, Yale UniversityDepartment of Microbial Pathogenesis, Yale University School of Medicine||Department of Comparative Medicine, Yale UniversityDepartment of Immunobiology, Yale University School of Medicine

10.1101/434027

基础医学分子生物学

Parker Michael T.,Gopinath Smita,Perez Corey E.,Crawford Jason M.,Iwasaki Akiko,Lindenbach Brett D.,Linehan Melissa M..Innate Immune Priming by cGAS as a Preparatory Countermeasure Against RNA Virus Infection[EB/OL].(2025-03-28)[2025-05-17].https://www.biorxiv.org/content/10.1101/434027.点此复制

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