title: Membrane Perturbation-Associated Ca2+ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry.
abstract: The type I interferon (IFN) response is an important aspect of innate antiviral defense, and the transcription factor IRF3 plays an important role in its induction. Membrane perturbation during fusion, a necessary step for enveloped virus particle entry, appears sufficient to induce transcription of a subset of IFN-stimulated genes (ISGs) in an IRF3-dependent, IFN-independent fashion. IRF3 is emerging as a central node in host cell stress responses, although it remains unclear how different forms of stress activate IRF3. Here, we investigated the minimum number of Sendai virus (SeV) and human cytomegalovirus (HCMV) particles required to activate IRF3 and trigger an antiviral response. We found that Ca(2+) signaling associated with membrane perturbation and recognition of incoming viral genomes by cytosolic nucleic acid receptors are required to activate IRF3 in response to fewer than 13 particles of SeV and 84 particles of HCMV per cell. Moreover, it appears that Ca(2+) signaling is important for activation of STING and IRF3 following HCMV particle entry, suggesting that Ca(2+) signaling sensitizes cells to recognize genomes within incoming virus particles. To our knowledge, this is the first evidence that cytosolic nucleic acid sensors recognize genomes within incoming virus particles prior to virus replication. These studies highlight the exquisite sensitivity of the cellular response to low-level stimuli and suggest that virus particle entry is sensed as a stress signal.
  
  
  OBJECTIVE
  The mechanism by which replicating viruses trigger IRF3 activation and type I IFN induction through the generation and accumulation of viral pathogen-associated molecular patterns has been well characterized. However, the mechanism by which enveloped virus particle entry mediates a stress response, leading to IRF3 activation and the IFN-independent response, remained elusive. Here, we find that Ca(2+) signaling associated with membrane perturbation appears to sensitize cells to recognize genomes within incoming virus particles. To our knowledge, this is the first study to show that cytosolic receptors recognize genomes within incoming virus particles prior to virus replication. These findings not only highlight the sensitivity of cellular responses to low-level virus particle stimulation, but provide important insights into how nonreplicating virus vectors or synthetic lipid-based carriers used as clinical delivery vehicles activate innate immune responses.
authors.affiliation: Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada.
authors.forename: David N
authors.initials: DN
authors.lastname: Hare
authors.affiliation: Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Collins
authors.affiliation: Firestone Institute for Respiratory Health, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Subhendu
authors.initials: S
authors.lastname: Mukherjee
authors.affiliation: Department of Immunology, Center for Innate Immunity and Immune Disease, School of Medicine, University of Washington, Seattle, Washington, USA.
authors.forename: Yueh-Ming
authors.identifier: http://orcid.org/0000-0001-6294-7273
authors.initials: YM
authors.lastname: Loo
authors.affiliation: Department of Immunology, Center for Innate Immunity and Immune Disease, School of Medicine, University of Washington, Seattle, Washington, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Gale
authors.affiliation: Firestone Institute for Respiratory Health, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Luke J
authors.initials: LJ
authors.lastname: Janssen
authors.affiliation: Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada mossk@mcmaster.ca.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Mossman
mesh_terms: D000818:Animals; D020013:Calcium Signaling; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology* / Q000502:physiology; D054884:Host-Pathogen Interactions; D006801:Humans; D007113:Immunity, Innate; D050838:Interferon Regulatory Factor-3 / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D029082:Sendai virus / Q000276:immunology* / Q000502:physiology; D053586:Virus Internalization
chemical_list: C494232:IRF3 protein, human; D050838:Interferon Regulatory Factor-3