Triggered by viral infection, the PRR-mediated immune response pathways are highly robust and complicated with the involvement of multiple, transiently assembled protein complexes that function in an interconnected and phenotypic manner. This complex assembly and diverse pathways demand new technology capable of systematically profiling those critical pathway components in more sensitive and precise approaches. We have previously demonstrated the unique strength of our discovery proteomics platform in identifying/profiling novel pathway constituents, including those dynamic protein-protein interactions (PPIs) and post-translational modifications (PTMs) involved in the regulation of the initiation, progression and termination of PRR-mediated immune signaling. We also showed that these results from the proteomic dissection of PRR-mediated pathways could guide and streamline in-depth functional analyses using conventional genetic, cell biology, and immunology approaches, thereby enlarging the scope of mechanistic understanding of the host immune response. Leveraging this extensive experience, the overall objective of this shared resource Core B is to enumerate the constituents of the subsequent signaling pathways caused by the interaction of innate immune PRRs with viruses. All quantitative proteomic analyses of PPIs and PTMs, the two major pathway operators, as well as virus-induced protein translocations, can be performed comparatively for infected subjects versus non-infected controls or at different infection stages in either global or subcellular compartments or in organelle-specific fashion. Various quantitative schemes including AACT/SILAC, ITRAQ, and label-free quantitation will be complimentarily used in our experiments for not only cell lines, but also primary cells or tissues. Additionally, we will be significantly involved in guiding sample preparation in all Projects, particularly those associated with newly developed techniques of proteomics. Results will generate multiple hypotheses (i.e. discovery of novel proteins, PPIs or PTMs) in both interconnected and efficient manners to guide concurrently the discovery and elucidation of novel biologies, pathways and mechanisms. Specifically by applying the state-of-the-art technology of quantitative proteomics, we will (1) dissect either global or pathway-specific protein-protein interaction networks (interactome) involved in the PRR-mediated immune response to viral infection, and (2) identify/profile global or interactome-specific post-translational modifications (PTMs) including phosphorylation, acetylation, and ubiquitination involved in the PRR-mediated immune response to viral infection.

Public Health Relevance

PRR-mediated intracellular signaling pathways play regulatory roles in the host immune response to viral infection. The Proteomics Pathway Discovery Core will use a varity of quantitative proteomics methodologies to identify and profile novel components of virus-inducible PRR-mediated signaling pathways. Results will guide and streamline new pathway characterization as defined by Projects 1-3 for revealing the immune mechanisms that lead to immunopathology versus a protective immune response during virus infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
1U19AI109965-01
Application #
8675630
Study Section
Special Emphasis Panel (ZAI1-ZL-I (J1))
Project Start
Project End
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
1
Fiscal Year
2014
Total Cost
$405,164
Indirect Cost
$138,275
Name
University of North Carolina Chapel Hill
Department
Type
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Hirai-Yuki, Asuka; Hensley, Lucinda; McGivern, David R et al. (2016) MAVS-dependent host species range and pathogenicity of human hepatitis A virus. Science 353:1541-1545
Erdoğan, Özgün; Xie, Ling; Wang, Li et al. (2016) Proteomic dissection of LPS-inducible, PHF8-dependent secretome reveals novel roles of PHF8 in TLR4-induced acute inflammation and T cell proliferation. Sci Rep 6:24833
Ma, Zhe; Damania, Blossom (2016) The cGAS-STING Defense Pathway and Its Counteraction by Viruses. Cell Host Microbe 19:150-8
Bhatt, Aadra Prashant; Wong, Jason P; Weinberg, Marc S et al. (2016) A viral kinase mimics S6 kinase to enhance cell proliferation. Proc Natl Acad Sci U S A 113:7876-81
Guo, Haitao; König, Renate; Deng, Meng et al. (2016) NLRX1 Sequesters STING to Negatively Regulate the Interferon Response, Thereby Facilitating the Replication of HIV-1 and DNA Viruses. Cell Host Microbe 19:515-28
Gunawardena, Harsha P; O'Brien, Jonathon; Wrobel, John A et al. (2016) QuantFusion: Novel Unified Methodology for Enhanced Coverage and Precision in Quantifying Global Proteomic Changes in Whole Tissues. Mol Cell Proteomics 15:740-51
Damania, Blossom (2016) A Virological Perspective on Cancer. PLoS Pathog 12:e1005326
Chatterjee, Srirupa; Basler, Christopher F; Amarasinghe, Gaya K et al. (2016) Molecular Mechanisms of Innate Immune Inhibition by Non-Segmented Negative-Sense RNA Viruses. J Mol Biol 428:3467-82
Giffin, Louise; West, John A; Damania, Blossom (2015) Kaposi's Sarcoma-Associated Herpesvirus Interleukin-6 Modulates Endothelial Cell Movement by Upregulating Cellular Genes Involved in Migration. MBio 6:e01499-15
Wang, Li; Xie, Ling; Ramachandran, Srinivas et al. (2015) Non-canonical Bromodomain within DNA-PKcs Promotes DNA Damage Response and Radioresistance through Recognizing an IR-Induced Acetyl-Lysine on H2AX. Chem Biol 22:849-61

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