2WHX

A second conformation of the NS3 protease-helicase from dengue virus

Summary for 2WHX

• Entry DOI: 10.2210/pdb2whx/pdb
• Related: 2JLQ 2JLR 2JLS 2JLU 2JLV 2JLW 2JLX 2JLY 2JLZ 2WZQ
• Descriptor: SERINE PROTEASE/NTPASE/HELICASE NS3, SERINE PROTEASE SUBUNIT NS2B, ADENOSINE-5'-DIPHOSPHATE, ... (6 entities in total)
• Functional Keywords: transcription, hydrolase, atp-binding, reticulum, nucleotidyltransferase, multifunctional enzyme, transcription regulation, ribonucleoprotein, rna-directed rna polymerase, rna replication, envelope protein
• Biological source: DENGUE VIRUS 4; More
• Cellular location: Capsid protein C: Virion (Potential). Peptide pr: Secreted (By similarity). Small envelope protein M: Virion membrane; Multi-pass membrane protein (By similarity). Envelope protein E: Virion membrane; Multi- pass membrane protein (By similarity). Non-structural protein 1: Secreted. Non-structural protein 2A-alpha: Host endoplasmic reticulum membrane; Multi-pass membrane protein (Potential). Non-structural protein 2A: Host endoplasmic reticulum membrane; Multi-pass membrane protein (Potential). Serine protease subunit NS2B: Host endoplasmic reticulum membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Serine protease NS3: Host endoplasmic reticulum membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Non-structural protein 4A: Host endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Non-structural protein 4B: Host endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). RNA-directed RNA polymerase NS5: Host endoplasmic reticulum membrane; Peripheral membrane protein; Cytoplasmic side (By similarity): Q2YHF0 Q2YHF0
• Total number of polymer chains: 2
• Total formula weight: 71602.90

Authors

Luo, D.,Lescar, J. (deposition date: 2009-05-07, release date: 2010-04-07, Last modification date: 2024-05-08)

Primary citation

Luo, D.,Wei, N.,Doan, D.N.,Paradkar, P.N.,Chong, Y.,Davidson, A.D.,Kotaka, M.,Lescar, J.,Vasudevan, S.G.
Flexibility between the Protease and Helicase Domains of the Dengue Virus Ns3 Protein Conferred by the Linker Region and its Functional Implications.
J.Biol.Chem., 285:18817-, 2010

PubMed 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 A of the soluble NS3 protein from DENV4 covalently attached to 18 residues of the NS2B cofactor region (NS2B(18)NS3) 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 approximately 161 degrees with respect to the helicase domain. We report this new crystal structure bound to ADP-Mn(2+) refined to a resolution of 2.2 A. The biological significance for interdomain flexibility conferred by the linker region was probed by either inserting a Gly residue between Glu(173) and Pro(174) or replacing Pro(174) with a Gly residue. Both mutations resulted in significantly lower ATPase and helicase activities. We next increased flexibility in the linker by introducing a Pro(176) 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.

PubMed: 20375022
DOI: 10.1074/JBC.M109.090936

Experimental method

X-RAY DIFFRACTION (2.2 Å)