Abstract
AbstractAlphaviruses such as Ross River virus (RRV), chikungunya virus, Sindbis virus (SINV), and Venezuelan equine encephalitis virus are mosquito-borne pathogens that can cause arthritis or encephalitis diseases. Nonstructural protein 4 (nsP4) of alphaviruses possesses RNA-dependent RNA polymerase (RdRp) activity essential for viral RNA replication. No 3D structure has been available for nsP4 of any alphaviruses despite its importance for understanding alphaviral RNA replication and for the design of antiviral drugs. Here, we report crystal structures of the RdRp domain of nsP4 from both RRV and SINV determined at resolutions of 2.6 and 1.9 Å. The structure of the alphavirus RdRp domain appears most closely related to RdRps from pestiviruses, noroviruses, and picornaviruses. Hydrogendeuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) methods, showed that in solution, nsP4 is highly dynamic with an intrinsically disordered N-terminal domain. Both full-length nsP4 and the RdRp domain were capable to catalyze RNA polymerization. Structure-guided mutagenesis using a trans-replicase system identified nsP4 regions critical for viral RNA replication.Key PointsCrystal structures of alphavirus nsP4 RNA polymerase domain from RRV and SINV.nsP4 protein is highly dynamic with an intrinsically disordered N-terminal domain.Optimized RNA elongation activity assay to facilitate antiviral discovery.
Publisher
Cold Spring Harbor Laboratory
Reference58 articles.
1. Global distribution and environmental suitability for chikungunya virus, 1952 to 2015;Eurosurveillance,2016
2. Lessler, J. , Chaisson, L.H. , Kucirka, L.M. , Bi, Q. , Grantz, K. , Salje, H. , Carcelen, A.C. , Ott, C.T. , Sheffield, J.S. and Ferguson, N.M. (2016) Assessing the global threat from Zika virus. Science, 353.
3. Analysis of coevolution in nonstructural proteins of chikungunya virus;Virol J,2016
4. Genome-Scale Phylogeny of the Alphavirus Genus Suggests a Marine Origin
Cited by
4 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献