Arenaviruses such as Lassa fever virus (LASV) can cause acute viral hemorrhagic fever disease in humans to which there is no vaccine or effective treatment. The mechanism of arenavirus replication in the infected cells is poorly understood, which has impeded the development of effective antivirals and vaccines. Arenaviral mRNA contains several nontemplated nucleotides that are believed to derive from the host cell messenger RNAs through the cap-snatching mechanism. To date, little is known about the specific protein(s) and the molecular mechanism that are involved in the cap-snatching process for arenaviruses. We have recently obtained a 3-D crystal structure of the first arenavirus protein, the LASV nucleoprotein NP, and identified several novel structural motifs that suggest NP as the first arenaviral protein involved in the cap-snatching process. We hypothesize that NP is an essential component of viral replication complex that functions to steal the 5 cap structures of cellular mRNA in order to initiate viral transcription. The main goal of this study, therefore, is to combine our strengths in molecular virology, structural modeling, and RNA/protein biochemistry and the availability of several novel viral reverse genetics systems in our laboratory to clearly define the molecular mechanism of cap snatching in arenaviruses. Specifically, we will characterize the cap-snatching function of the LASV NP protein in vitro by biochemical analyses (Aim 1) and in vivo by several novel and functional cell-based assays and infectious virus systems (Aim 2). Lastly, we wish to characterize the molecular interactions between the viral NP and L proteins within the replication complex in mediating and/or regulating the cap-snatching process (Aim 3). Taken together, these studies will reveal for the first time a novel molecular mechanism of the cap-snatching step for arenavirus and its biological significance in transcribing the viral genome in vitro and in vivo. These studies will shed important lights on arenaviral RNA transcription that may offer opportunities for development of effective therapeutics and/or vaccines against lethal arenaviral hemorrhagic fever infection in humans.

Public Health Relevance

Lassa fever virus infection can lead to severe and sometime lethal hemorrhagic fever diseases in humans, for which there is no vaccine and effective antivirals. Little has been known of the molecular mechanism by which arenavirus steals the cap structure from host mRNAs (cap snatching) to initiate viral RNA transcription. Based on the first 3D-crystal structure of the LASV nucleoprotein (NP) that we have recently obtained, we hypothesize that arenavirus NP protein is an essential component required for cap snatching, which suggests a distinct mechanism from the well-characterized influenza virus system. In this proposal, we will clearly characterize the novel mechanism of cap snatching process in arenavirus. Results from these studies may lead to identification of antiviral candidates and the development of effective therapeutics and vaccines against some deadly arenavirus-induced hemorrhagic diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
High Priority, Short Term Project Award (R56)
Project #
1R56AI091805-01
Application #
8293447
Study Section
Virology - A Study Section (VIRA)
Program Officer
Repik, Patricia M
Project Start
2011-08-01
Project End
2011-10-30
Budget Start
2011-08-01
Budget End
2011-10-30
Support Year
1
Fiscal Year
2011
Total Cost
$23,467
Indirect Cost
Name
Emory University
Department
Pathology
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Qi, Xiaoxuan; Wang, Wenjian; Dong, Haohao et al. (2018) Expression and X-Ray Structural Determination of the Nucleoprotein of Lassa Fever Virus. Methods Mol Biol 1604:179-188
Dhanwani, Rekha; Huang, Qinfeng; Lan, Shuiyun et al. (2018) Establishment of Bisegmented and Trisegmented Reverse Genetics Systems to Generate Recombinant Pichindé Viruses. Methods Mol Biol 1604:247-253
Huang, Qinfeng; Shao, Junjie; Liang, Yuying et al. (2018) Assays to Demonstrate the Roles of Arenaviral Nucleoproteins (NPs) in Viral RNA Synthesis and in Suppressing Type I Interferon. Methods Mol Biol 1604:189-200
Shao, Junjie; Liang, Yuying; Ly, Hinh (2018) Roles of Arenavirus Z Protein in Mediating Virion Budding, Viral Transcription-Inhibition and Interferon-Beta Suppression. Methods Mol Biol 1604:217-227
Meyer, Bjoern; Ly, Hinh (2016) Inhibition of Innate Immune Responses Is Key to Pathogenesis by Arenaviruses. J Virol 90:3810-3818
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Wallat, Gregor D; Huang, Qinfeng; Wang, Wenjian et al. (2014) High-resolution structure of the N-terminal endonuclease domain of the Lassa virus L polymerase in complex with magnesium ions. PLoS One 9:e87577
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McLay, Lisa; Ansari, Aftab; Liang, Yuying et al. (2013) Targeting virulence mechanisms for the prevention and therapy of arenaviral hemorrhagic fever. Antiviral Res 97:81-92
McLay, Lisa; Lan, Shuiyun; Ansari, Aftab et al. (2013) Identification of virulence determinants within the L genomic segment of the pichinde arenavirus. J Virol 87:6635-43

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