Please use this identifier to cite or link to this item: https://doi.org/10.1074/jbc.M109.090936
Title: Flexibility between the protease and helicase domains of the dengue virus NS3 protein conferred by the linker region and its functional implications
Authors: Luo, D.
Wei, N.
Doan, D.N.
Paradkar, P.N.
Chong, Y.
Davidson, A.D.
Kotaka, M.
Lescar, J.
Vasudevan, S.G. 
Issue Date: 11-Jun-2010
Source: Luo, D., Wei, N., Doan, D.N., Paradkar, P.N., Chong, Y., Davidson, A.D., Kotaka, M., Lescar, J., Vasudevan, S.G. (2010-06-11). Flexibility between the protease and helicase domains of the dengue virus NS3 protein conferred by the linker region and its functional implications. Journal of Biological Chemistry 285 (24) : 18817-18827. ScholarBank@NUS Repository. https://doi.org/10.1074/jbc.M109.090936
Abstract: The dengue virus (DENV) NS3 protein is essential for viral polyprotein processing and RNA replication. It contains an N-terminal serine protease region (residues 1-168) joined to an RNA helicase (residues 180-618) by an 11-amino acid linker (169-179). The structure at 3.15 Å of the soluble NS3 protein from DENV4 covalently attached to 18 residues of the NS2B cofactor region (NS2B18NS3) revealed an elongated molecule with the protease domain abutting subdomains I and II of the helicase (Luo, D., Xu, T., Hunke, C., Grüber, G., Vasudevan, S. G., and Lescar, J. (2008) J. Virol. 82, 173-183). Unexpectedly, using similar crystal growth conditions, we observed an alternative conformation where the protease domain has rotated by ∼161° with respect to the helicase domain. We report this new crystal structure bound to ADP-Mn2+ refined to a resolution of 2.2 Å. The biological significance for interdomain flexibility conferred by the linker region was probed by either inserting a Gly residue between Glu173 and Pro 174 or replacing Pro174 with a Gly residue. Both mutations resulted in significantly lower ATPase and helicase activities. We next increased flexibility in the linker by introducing a Pro176 to Gly mutation in a DENV2 replicon system. A 70% reduction in luciferase reporter signal and a similar reduction in the level of viral RNA synthesis were observed. Our results indicate that the linker region has evolved to an optimum length to confer flexibility to the NS3 protein that is required both for polyprotein processing and RNA replication. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.
Source Title: Journal of Biological Chemistry
URI: http://scholarbank.nus.edu.sg/handle/10635/110086
ISSN: 00219258
DOI: 10.1074/jbc.M109.090936
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