Project Overview for Structural and Functional Studies of Ebola Virus RNA synthesis Ebolaviruses (EBOVs) are non-segmented negative-strand RNA viruses (NNSVs) and Category A Priority biodefense pathogens that cause frequent lethal hemorrhagic fever, including the devastating and ongoing outbreak in West Africa. Approved anti-EBOV therapeutics are lacking. EBOV replicates with high efficiency in vivo while effectively evading host innate antiviral immunity. The unrestrained replication and associated inflammation leads to death. A complete understanding of EBOV pathogenesis therefore requires understanding how the viral RNA dependent RNA polymerase (RDRP) complex interacts with host factors. The overarching goal of this program project proposal is to understand the mechanisms of RDRP function by defining the interactions among its components and with the host proteome and by defining signals that regulate its function. Toward this goal Project 1 will identify cis- and trans-acting factors that impact EBOV RDRP activity. Project 2 will define a structural basis for the EBOV RDRP complex, identify regulatory mechanisms, and determine species and strain specificities, and Project 3 will use genome-wide RNAi and proteomic strategies to identify host factors that modulate RDRP function. Project 1 provides functional context into which a subset of Project 3 ?hits? can be interpreted, while Project 3 will likely identify host factors relevant to the findings of Project 1. Project 2 will provide a structural framework that will be used to integrate findings from both Projects 1 and 3. Together, these studies will provide mechanistic insights into how protein-protein and protein-vRNA interactions control EBOV RDRP function. Research projects are supported by an Administrative Core, a Protein Production and Protein Interaction Core, and critically, a Biosafety Level 4 (BSL4) Core. Core B will generate unique reagents, including monoclonal and synthetic antibodies. Core C (BSL4) will provide unique capabilities to obtain materials from and perform tests using wild type and mutant EBOVs as well as pathogenesis evaluation. The work will be performed by highly productive and collaborative investigators with expertise in every aspect of the proposed studies, including biochemistry, EBOV pathogenesis, high throughput screening and data analysis, immunology, proteomics, structural biology, and virology. These studies will provide unprecedented insights into the components of the EBOV RDRP and in its interaction with the host as well as species and strain differences that affect the RDRP complex. We also expect to identify specific viral and host factor interactions as novel therapeutic targets.
Project Overview for Structural and Functional Studies of Ebola Virus RNA synthesis Ebola viruses are important emerging and biodefense pathogens for which approved therapeutics are lacking. The characterization of the viral RNA polymerase complex and its interactions with host proteins by this interdisciplinary project will shed light on viral replication and identify new therapeutic targets.
|Johnson, Britney; McConnell, Patrick; Kozlov, Alex G et al. (2018) Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange. Cell Rep 23:2795-2804|
|Holze, Cathleen; Michaudel, Chloé; Mackowiak, Claire et al. (2018) Oxeiptosis, a ROS-induced caspase-independent apoptosis-like cell-death pathway. Nat Immunol 19:130-140|
|Warfield, Kelly L; Howell, Katie A; Vu, Hong et al. (2018) Role of Antibodies in Protection Against Ebola Virus in Nonhuman Primates Immunized With Three Vaccine Platforms. J Infect Dis 218:S553-S564|
|Su, Zhaoming; Wu, Chao; Shi, Liuqing et al. (2018) Electron Cryo-microscopy Structure of Ebola Virus Nucleoprotein Reveals a Mechanism for Nucleocapsid-like Assembly. Cell 172:966-978.e12|
|Knoverek, Catherine R; Amarasinghe, Gaya K; Bowman, Gregory R (2018) Advanced Methods for Accessing Protein Shape-Shifting Present New Therapeutic Opportunities. Trends Biochem Sci :|
|Johnson, Britney; VanBlargan, Laura A; Xu, Wei et al. (2018) Human IFIT3 Modulates IFIT1 RNA Binding Specificity and Protein Stability. Immunity 48:487-499.e5|
|Klein, Roger D; Shu, Qin; Cusumano, Zachary T et al. (2018) Structure-Function Analysis of the Curli Accessory Protein CsgE Defines Surfaces Essential for Coordinating Amyloid Fiber Formation. MBio 9:|
|Hung, Putzer J; Johnson, Britney; Chen, Bo-Ruei et al. (2018) MRI Is a DNA Damage Response Adaptor during Classical Non-homologous End Joining. Mol Cell 71:332-342.e8|
|Johnston, Adam B; Hilton, Denise M; McConnell, Patrick et al. (2018) A novel mode of capping protein-regulation by twinfilin. Elife 7:|
|Dashti, Hesam; Wedell, Jonathan R; Westler, William M et al. (2018) Applications of Parametrized NMR Spin Systems of Small Molecules. Anal Chem 90:10646-10649|
Showing the most recent 10 out of 17 publications