title: Inhibition of interferon gene activation by death-effector domain-containing proteins from the molluscum contagiosum virus.
abstract: Apoptosis, NF-κB activation, and IRF3 activation are a triad of intrinsic immune responses that play crucial roles in the pathogenesis of infectious diseases, cancer, and autoimmunity. FLIPs are a family of viral and cellular proteins initially found to inhibit apoptosis and more recently to either up- or down-regulate NF-κB. As such, a broad role for FLIPs in disease regulation is postulated, but exactly how a FLIP performs such multifunctional roles remains to be established. Here we examine FLIPs (MC159 and MC160) encoded by the molluscum contagiosum virus, a dermatotropic poxvirus causing skin infections common in children and immunocompromised individuals, to better understand their roles in viral pathogenesis. While studying their molecular mechanisms responsible for NF-κB inhibition, we discovered that each protein inhibited IRF3-controlled luciferase activity, identifying a unique function for FLIPs. MC159 and MC160 each inhibited TBK1 phosphorylation, confirming this unique function. Surprisingly, MC159 coimmunoprecipitated with TBK1 and IKKε but MC160 did not, suggesting that these homologs use distinct molecular mechanisms to inhibit IRF3 activation. Equally surprising was the finding that the FLIP regions necessary for TBK1 inhibition were distinct from those MC159 or MC160 regions previously defined to inhibit NF-κB or apoptosis. These data reveal previously unappreciated complexities of FLIPs, and that subtle differences within the conserved regions of FLIPs possess distinct molecular and structural fingerprints that define crucial differences in biological activities. A future comparison of mechanistic differences between viral FLIP proteins can provide new means of precisely manipulating distinct aspects of intrinsic immune responses.
authors.affiliation: Department of Microbiology, College of Medicine, University of Illinois, Urbana, IL 61801.
authors.forename: Crystal M H
authors.initials: CM
authors.lastname: Randall
authors.forename: Sunetra
authors.initials: S
authors.lastname: Biswas
authors.forename: Catherine V
authors.initials: CV
authors.lastname: Selen
authors.forename: Joanna L
authors.initials: JL
authors.lastname: Shisler
mesh_terms: D057809:HEK293 Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D015151:Immunoblotting; D047468:Immunoprecipitation; D016899:Interferon-beta / Q000037:antagonists & inhibitors*; D008156:Luciferases / Q000037:antagonists & inhibitors; D008976:Molluscum Contagiosum / Q000276:immunology*; D008977:Molluscum contagiosum virus / Q000235:genetics* / Q000276:immunology; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D016328:NF-kappa B; D014764:Viral Proteins; C509979:viral FLIP protein, Molluscum contagiosum virus; D016899:Interferon-beta; D008156:Luciferases; D017346:Protein Serine-Threonine Kinases; C403191:TBK1 protein, human; D051550:I-kappa B Kinase
keywords: host–pathogen interactions; immune evasion