Please use this identifier to cite or link to this item:
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
Citation: 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.
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
ISSN: 00219258
DOI: 10.1074/jbc.M109.090936
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.


checked on Jan 15, 2020


checked on Jan 8, 2020

Page view(s)

checked on Jan 18, 2020

Google ScholarTM



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.