title: An ITAM in a nonenveloped virus regulates activation of NF-κB, induction of beta interferon, and viral spread.
abstract: Immunoreceptor tyrosine-based activation motifs (ITAMs) are signaling domains located within the cytoplasmic tails of many transmembrane receptors and associated adaptor proteins that mediate immune cell activation. ITAMs also have been identified in the cytoplasmic tails of some enveloped virus glycoproteins. Here, we identified ITAM sequences in three mammalian reovirus proteins: μ2, σ2, and λ2. We demonstrate for the first time that μ2 is phosphorylated, contains a functional ITAM, and activates NF-κB. Specifically, μ2 and μNS recruit the ITAM-signaling intermediate Syk to cytoplasmic viral factories and this recruitment requires the μ2 ITAM. Moreover, both the μ2 ITAM and Syk are required for maximal μ2 activation of NF-κB. A mutant virus lacking the μ2 ITAM activates NF-κB less efficiently and induces lower levels of the downstream antiviral cytokine beta interferon (IFN-β) than does wild-type virus despite similar replication. Notably, the consequences of these μ2 ITAM effects are cell type specific. In fibroblasts where NF-κB is required for reovirus-induced apoptosis, the μ2 ITAM is advantageous for viral spread and enhances viral fitness. Conversely, in cardiac myocytes where the IFN response is critical for antiviral protection and NF-κB is not required for apoptosis, the μ2 ITAM stimulates cellular defense mechanisms and diminishes viral fitness. Together, these results suggest that the cell type-specific effect of the μ2 ITAM on viral spread reflects the cell type-specific effects of NF-κB and IFN-β. This first demonstration of a functional ITAM in a nonenveloped virus presents a new mechanism for viral ITAM-mediated signaling with likely organ-specific consequences in the host.
authors.affiliation: Department of Molecular Biomedical Sciences and Center for Comparative Medicine and Translational Research, North Carolina State University, Raleigh, North Carolina, USA.
authors.forename: Rachael E
authors.initials: RE
authors.lastname: Stebbing
authors.forename: Susan C
authors.initials: SC
authors.lastname: Irvin
authors.forename: Efraín E
authors.initials: EE
authors.lastname: Rivera-Serrano
authors.forename: Karl W
authors.initials: KW
authors.lastname: Boehme
authors.forename: Mine
authors.initials: M
authors.lastname: Ikizler
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Yoder
authors.forename: Terence S
authors.initials: TS
authors.lastname: Dermody
authors.forename: Barbara
authors.initials: B
authors.lastname: Sherry
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D061625:Immunoreceptor Tyrosine-Based Activation Motif; D016899:Interferon-beta / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000821:virology; D016328:NF-kappa B / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D012087:Reoviridae / Q000472:pathogenicity / Q000502:physiology*; D016415:Sequence Alignment; D000072377:Syk Kinase; D014443:Tyrosine / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*; D056189:Viral Tropism
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D016328:NF-kappa B; D014764:Viral Proteins; D014443:Tyrosine; D016899:Interferon-beta; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C000606897:Syk protein, mouse