@article{ff27a4e4e84a4a0dba6c49b1335d5cf9,
title = "In silico derived small molecules bind the filovirus VP35 protein and inhibit its polymerase cofactor activity",
abstract = "The Ebola virus (EBOV) genome only encodes a single viral polypeptide with enzymatic activity, the viral large (L) RNA-dependent RNA polymerase protein. However, currently, there is limited information about the L protein, which has hampered the development of antivirals. Therefore, antifiloviral therapeutic efforts must include additional targets such as protein-protein interfaces. Viral protein 35 (VP35) is multifunctional and plays important roles in viral pathogenesis, including viral mRNA synthesis and replication of the negative-sense RNA viral genome. Previous studies revealed that mutation of key basic residues within the VP35 interferon inhibitory domain (IID) results in significant EBOV attenuation, both in vitro and in vivo. In the current study, we use an experimental pipeline that includes structure-based in silico screening and biochemical and structural characterization, along with medicinal chemistry, to identify and characterize small molecules that target a binding pocket within VP35. NMR mapping experiments and high-resolution x-ray crystal structures show that select small molecules bind to a region of VP35 IID that is important for replication complex formation through interactions with the viral nucleoprotein (NP). We also tested select compounds for their ability to inhibit VP35 IID-NP interactions in vitro as well as VP35 function in a minigenome assay and EBOV replication. These results confirm the ability of compounds identified in this study to inhibit VP35-NP interactions in vitro and to impair viral replication in cell-based assays. These studies provide an initial framework to guide development of antifiloviral compounds against filoviral VP35 proteins.",
keywords = "VP35, antivirals, filoviral inhibitors, in silico drug discovery",
author = "Brown, {Craig S.} and Lee, {Michael S.} and Leung, {Daisy W.} and Tianjiao Wang and Wei Xu and Priya Luthra and Manu Anantpadma and Shabman, {Reed S.} and Melito, {Lisa M.} and Macmillan, {Karen S.} and Borek, {Dominika M.} and Zbyszek Otwinowski and Parameshwaran Ramanan and Stubbs, {Alisha J.} and Peterson, {Dayna S.} and Binning, {Jennifer M.} and Marco Tonelli and Olson, {Mark A.} and Davey, {Robert A.} and Ready, {Joseph M.} and Basler, {Christopher F.} and Amarasinghe, {Gaya K.}",
note = "Funding Information: We thank Drs. S. Ginell, N. Duke, F. Rotella, K. Lazarski, and J. Lazarz at Argonne National Laboratory Structural Biology Center Beamlines (SBC). SBC is supported by the U.S. Department of Energy under contract DE-AC02-06CH11357. D.M.B. and Z.O. are supported in part by the Center for Structural Genomics of Infectious Diseases under the NIAID/NIH/DHS Contracts HHSN272200700058C and HHSN272201200026C Anderson (PI). This work is supported in part by National Institutes of Health grants ( R01AI081914 to G.K.A. and R01AI059536 , R56AI089547 , R56AI093786 , and R21AI097568 to C.F.B.), a Midwest Regional Center of Excellence Developmental Grant [ U54AI057160 -Virgin (PI) to G.K.A.], and the Welch Foundation ( I-1612 to J.M.R.). DoD Defense Threat Reduction Agency Grant 4.10011_07_RD_B to M. A. O. This study made use of the National Magnetic Resonance Facility at Madison, which is supported by National Institutes of Health grants P41RR02301 (Biomedical Research Technology Program, National Center for Research Resources) and P41GM66326 (National Institute of General Medical Sciences). Equipment in the facility was purchased with funds from the University of Wisconsin, the National Institutes of Health ( P41GM66326 , P41RR02301 , RR02781 , and RR08438 ), the National Science Foundation ( DMB-8415048 , OIA-9977486 , and BIR-9214394 ), and the U.S. Department of Agriculture. ",
year = "2014",
month = may,
day = "15",
doi = "10.1016/j.jmb.2014.01.010",
language = "English (US)",
volume = "426",
pages = "2045--2058",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "10",
}