dc.contributor.authorZhao, Yongqian
dc.contributor.authorSoh, Tingjin Sherryl
dc.contributor.authorZheng, Jie
dc.contributor.authorChan, Kitti Wing Ki
dc.contributor.authorPhoo, Wint Wint
dc.contributor.authorLee, Chin Chin
dc.contributor.authorTay, Moon Y. F.
dc.contributor.authorSwaminathan, Kunchithapadam
dc.contributor.authorCornvik, Tobias Carl
dc.contributor.authorLim, Siew Pheng
dc.contributor.authorShi, Pei-Yong
dc.contributor.authorLescar, Julien
dc.contributor.authorVasudevan, Subhash G.
dc.contributor.authorLuo, Dahai
dc.contributor.editorRey, Félix A.*
dc.date.accessioned2015-05-18T02:22:50Z
dc.date.available2015-05-18T02:22:50Z
dc.date.copyright2015en_US
dc.date.issued2015
dc.identifier.citationZhao, Y., Soh, T. S., Zheng, J., Chan, K. W. K., Phoo, W. W., Lee, C. C., et al. (2015). A crystal structure of the dengue virus NS5 protein reveals a novel inter-domain interface essential for protein flexibility and virus replication. PLOS pathogens, 11(3).en_US
dc.identifier.issn1553-7374en_US
dc.identifier.urihttp://hdl.handle.net/10220/25565
dc.description.abstractFlavivirus RNA replication occurs within a replication complex (RC) that assembles on ER membranes and comprises both non-structural (NS) viral proteins and host cofactors. As the largest protein component within the flavivirus RC, NS5 plays key enzymatic roles through its N-terminal methyltransferase (MTase) and C-terminal RNA-dependent-RNA polymerase (RdRp) domains, and constitutes a major target for antivirals. We determined a crystal structure of the full-length NS5 protein from Dengue virus serotype 3 (DENV3) at a resolution of 2.3 Å in the presence of bound SAH and GTP. Although the overall molecular shape of NS5 from DENV3 resembles that of NS5 from Japanese Encephalitis Virus (JEV), the relative orientation between the MTase and RdRp domains differs between the two structures, providing direct evidence for the existence of a set of discrete stable molecular conformations that may be required for its function. While the inter-domain region is mostly disordered in NS5 from JEV, the NS5 structure from DENV3 reveals a well-ordered linker region comprising a short 310 helix that may act as a swivel. Solution Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) analysis reveals an increased mobility of the thumb subdomain of RdRp in the context of the full length NS5 protein which correlates well with the analysis of the crystallographic temperature factors. Site-directed mutagenesis targeting the mostly polar interface between the MTase and RdRp domains identified several evolutionarily conserved residues that are important for viral replication, suggesting that inter-domain cross-talk in NS5 regulates virus replication. Collectively, a picture for the molecular origin of NS5 flexibility is emerging with profound implications for flavivirus replication and for the development of therapeutics targeting NS5.en_US
dc.description.sponsorshipNMRC (Natl Medical Research Council, S’pore)
dc.format.extent27 p.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesPLOS pathogensen_US
dc.rights© 2015 Zhao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.subjectDRNTU::Science::Biological sciences::Microbiology::Bacteria
dc.titleA crystal structure of the dengue virus NS5 protein reveals a novel inter-domain interface essential for protein flexibility and virus replicationen_US
dc.typeJournal Article
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.contributor.schoolLee Kong Chian School of Medicine (LKCMedicine)
dc.identifier.doihttp://dx.doi.org/10.1371/journal.ppat.1004682
dc.description.versionPublished versionen_US


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