Many viruses activate the cellular NF-kappa B transcription factor as a method to express their own viral genes. However, NF-kappa B is dangerous as it induces expression of many anti-viral immune and inflammatory molecules. Thus, many viruses also produce proteins to inhibit NF-kappa B activation. Obviously, the timing, degree and pathways for NF-kappa B activation must be precisely regulated by viruses to ensure virus gene expression and immune evasion. Poxvirus infections both induce and inhibit NF-kappa B activation. However, very few individual genes have been identified as being responsible for these global effects. Even less clear is how poxviruses regulate NF-kappa B during infection to balance its beneficial and detrimental effects. Our working hypothesis is that vaccinia contains as yet undiscovered NF-kappa B inhibitors and activators, with inhibitory proteins responsible for dampening immune responses as a mechanism for virulence and activating proteins important for virus replication. Using a genetic approach, we have determined a 5.2 kilobase region of VV DNA that inhibits MVA-induced NF-kappa B activation. Known poxviral NF-kappa B inhibitors are not present in this 5.2 kb region, suggesting that VV genes are bona fide NF-kappa B inhibitors. To determine the novel NF-kappa B inhibitor(s), individual genes from this 5.2 kb region (namely the M1L, M2L and K1L genes) will be cloned and expressed independently in recombinant MVA viruses to identify NF-kappa B inhibitory activity. Using this approach, we have already identified the K1L and M2L products as NF-kappa B inhibitors (Specific Aim1). Defining the molecular mechanism used by K1L or M2L will give insight into each protein's function during infection (Specific Aim 2). Finally, the mechanism MVA uses to induce NF-kappa B activation will be defined, mapping the pathways MVA utilizes to induce NF-kappa B activation and the viral proteins responsible (Specific Aim 3). Results obtained here may prove useful in devising novel strategies to overcome pathogenic effects of vaccinia, with implications for enhancing safety and efficacy of a smallpox vaccine.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI055530-03
Application #
7177489
Study Section
Virology - A Study Section (VIRA)
Program Officer
Challberg, Mark D
Project Start
2005-02-01
Project End
2010-01-31
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
3
Fiscal Year
2007
Total Cost
$334,837
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Gates, Lauren T; Shisler, Joanna L (2016) cFLIPL Interrupts IRF3-CBP-DNA Interactions To Inhibit IRF3-Driven Transcription. J Immunol 197:923-33
Randall, Crystal M H; Jokela, Janet A; Shisler, Joanna L (2012) The MC159 protein from the molluscum contagiosum poxvirus inhibits NF-ýýB activation by interacting with the IýýB kinase complex. J Immunol 188:2371-9
Martin, Stefani; Harris, Daniel T; Shisler, Joanna (2012) The C11R gene, which encodes the vaccinia virus growth factor, is partially responsible for MVA-induced NF-ýýB and ERK2 activation. J Virol 86:9629-39
Willis, Kristen L; Langland, Jeffrey O; Shisler, Joanna L (2011) Viral double-stranded RNAs from vaccinia virus early or intermediate gene transcripts possess PKR activating function, resulting in NF-kappaB activation, when the K1 protein is absent or mutated. J Biol Chem 286:7765-78
Martin, Stefani; Shisler, Joanna L (2009) Early viral protein synthesis is necessary for NF-kappaB activation in modified vaccinia Ankara (MVA)-infected 293 T fibroblast cells. Virology 390:298-306
Nichols, Daniel Brian; Shisler, Joanna L (2009) Poxvirus MC160 protein utilizes multiple mechanisms to inhibit NF-kappaB activation mediated via components of the tumor necrosis factor receptor 1 signal transduction pathway. J Virol 83:3162-74
Willis, Kristen L; Patel, Samir; Xiang, Yan et al. (2009) The effect of the vaccinia K1 protein on the PKR-eIF2alpha pathway in RK13 and HeLa cells. Virology 394:73-81
Hinthong, Olivia; Jin, Xiao-Lu; Shisler, Joanna L (2008) Characterization of wild-type and mutant vaccinia virus M2L proteins'abilities to localize to the endoplasmic reticulum and to inhibit NF-kappaB activation during infection. Virology 373:248-62
Nichols, Daniel Brian; Shisler, Joanna L (2006) The MC160 protein expressed by the dermatotropic poxvirus molluscum contagiosum virus prevents tumor necrosis factor alpha-induced NF-kappaB activation via inhibition of I kappa kinase complex formation. J Virol 80:578-86
Gedey, Roderick; Jin, Xiao-Lu; Hinthong, Olivia et al. (2006) Poxviral regulation of the host NF-kappaB response: the vaccinia virus M2L protein inhibits induction of NF-kappaB activation via an ERK2 pathway in virus-infected human embryonic kidney cells. J Virol 80:8676-85

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