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Title: Multiple Enzyme Activities of Flavivirus Proteins
Authors: R. Padmanabhan
N. Mueller
E. Reichert
C. Yon
T. Teramoto
Y. Kono
R. Takhampunya
S. Ubol
N. Pattabiraman
B. Falgout
V. K. Ganesh
K. Murthy
Georgetown University School of Medicine
Mahidol University
Food and Drug Administration
University of Alabama
Keywords: Immunology and Microbiology
Issue Date: 7-Oct-2008
Citation: New Treatment Strategies for Dengue and Other Flaviviral Diseases. (2008), 74-86
Abstract: © 2006 Novartis Foundation. All Rights Reserved. Dengue viruses (DENV) have 5′-capped RNA genomes of (+) polarity and encode a single polyprotein precursor that is processed into mature viral proteins. NS2B, NS3 and NS5 proteins catalyse/activate enzyme activities that are required for key processes in the virus life cycle. The heterodimeric NS2B/NS3 is a serine protease required for processing. Using a high-throughput protease assay, we screened a small molecule chemical library and identified ∼200 compounds having ≥50% inhibition. Moreover, NS3 exhibits RNA-stimulated NTPase, RNA helicase and the 5′-RNA triphosphatase activities. The NTPase and the 5′-RTPase activities of NS3 are stimulated by interaction with NS5. Moreover, the conserved, positively charged motif in DENV-2 NS3, 184RKRK, is required for RNA binding and modulates the RNA-dependent enzyme activities of NS3. To study viral replication, a variety of methods are used such as the in vitro RNAdependent RNA polymerase assays that utilize lysates from DENV-2-infected mosquito- or mammalian cells or the purified NS5 along with exogenous short subgenomic viral RNAs or the replicative intracellular membrane-bound viral RNAs as templates. In addition, a cell-based DENV-2 replicon RNA encoding a luciferase reporter is also used to examine the role of cis-acting elements within the 3′ UTR and the RKRK motif in viral replication.
Appears in Collections:Scopus 2006-2010

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