Project 2: Structure and function of poxvirus immune evasion domains. In this project we will explore the functional and mechanistic attributes of a large family of sequence diverse proteins encoded by the zoonotic orthopoxvirus cowpox. We hypothesize that ten proteins In cowpox share a similar structural scaffolding that we have termed the poxvirus immune evasion (PIE) domain. Certain PIE proteins are known to sabotage host Immune surveillance and effector functions, and our goal Is to systematically determine the ligands and receptors for all ten PIE proteins encoded by the Brighton Red strain of cowpox. These efforts will be complemented by crystallographic investigations and quantitative binding assays to determine PIE protein structure and specificity, and structure-based engineering to develop variants with altered ligand/receptor Interactions. We will also undertake extensive In vitro functional studies of the PlEs, evaluating their mechanisms of action In the context of well-established Immunological assays. Our investigations will also be considered In the broader context of the battlefield of viral infection and host defense. In close collaboration with the Yokoyama and Fruh labs we will evaluate the pathogenesis of cowpox with either selective PIE proteins knocked out or their ligand/receptor binding determinants altered. The proposed experiments will result In a significantly improved functional annotation of cowpox and yield important clues about host-pathogen Interactions that are likely to benefit both the scientific and clinical communities.
A major bottleneck In understanding pathogenesis of Infectious disease viruses has been elucidating the entire spectrum of immune evasion strategies they deploy to ensure survival in their targeted hosts. We believe that successful completion of this proposal will expand the current repertoire of known poxvirus immune evasion strategies and may uncover novel host mechanisms that control infection and spread.
|Gilchuk, Iuliia; Gilchuk, Pavlo; Sapparapu, Gopal et al. (2016) Cross-Neutralizing and Protective Human Antibody Specificities to Poxvirus Infections. Cell 167:684-694.e9|
|Ghasemi, Reza; Lazear, Eric; Wang, Xiaoli et al. (2016) Selective targeting of IL-2 to NKG2D bearing cells for improved immunotherapy. Nat Commun 7:12878|
|Lubman, Olga Y; Fremont, Daved H (2016) Parallel Evolution of Chemokine Binding by Structurally Related Herpesvirus Decoy Receptors. Structure 24:57-69|
|Nelson, Christopher A; Epperson, Megan L; Singh, Sukrit et al. (2015) Structural Conservation and Functional Diversity of the Poxvirus Immune Evasion (PIE) Domain Superfamily. Viruses 7:4878-98|
|Nelson, Christopher A; Lee, Chung A; Fremont, Daved H (2014) Oxidative refolding from inclusion bodies. Methods Mol Biol 1140:145-57|
|Alzhanova, Dina; Hammarlund, Erika; Reed, Jason et al. (2014) T cell inactivation by poxviral B22 family proteins increases viral virulence. PLoS Pathog 10:e1004123|
|Luteijn, Rutger D; Hoelen, Hanneke; Kruse, Elisabeth et al. (2014) Cowpox virus protein CPXV012 eludes CTLs by blocking ATP binding to TAP. J Immunol 193:1578-89|
|Nelson, Christopher A; McCoy, William H; Fremont, Daved H (2014) Eukaryotic expression systems for structural studies. Methods Mol Biol 1140:107-16|