title: Structural features of Zika virus non-structural proteins 3 and -5 and its individual domains in solution as well as insights into NS3 inhibition.
abstract: Zika virus (ZIKV) has emerged as a pathogen of major health concern. The virus relies on its non-structural protein 5 (NS5) including a methyl-transferase (MTase) and a RNA-dependent RNA polymerase (RdRp) for capping and synthesis of the viral RNA and the nonstructural protein 3 (NS3) with its protease and helicase domain for polyprotein possessing, unwinding dsRNA proceeding replication, and NTPase/RTPase activities. In this study we present for the first time insights into the overall structure of the entire French Polynesia ZIKV NS3 in solution. The protein is elongated and flexible in solution. Solution studies of the individual protease- and helicase domains show the compactness of the two monomeric enzymes as well as the contribution of the 10-residues linker region to the flexibility of the entire NS3. We show also the solution X-ray scattering data of the French Polynesia ZIKV NS5, which is dimeric in solution and switches to oligomers in a concentration-dependent manner. The solution shapes of the MTase and RdRp domains are described. The dimer arrangement of ZIKV NS5 is discussed in terms of its importance for MTase-RdRp communication and concerted interaction with its flexible and monomeric counterpart NS3 during viral replication and capping. The comparison of ZIKV NS3 and -NS5 solution data with the related DENV nonstructural proteins shed light into the similarities and diversities of these classes of enzymes. Finally, the effect of ATPase inhibitors to the enzymatic active ZIKV NS3 and the individual helicase are provided.
authors.affiliation: Nanyang Technological University, School of Biological Sciences, 60 Nanyang Drive, 637551, Singapore.
authors.forename: Wuan Geok
authors.initials: WG
authors.lastname: Saw
authors.affiliation: Nanyang Technological University, School of Biological Sciences, 60 Nanyang Drive, 637551, Singapore.
authors.forename: Ankita
authors.initials: A
authors.lastname: Pan
authors.affiliation: Nanyang Technological University, School of Biological Sciences, 60 Nanyang Drive, 637551, Singapore.
authors.forename: Malathy Sony
authors.initials: MS
authors.lastname: Subramanian Manimekalai
authors.affiliation: Nanyang Technological University, School of Biological Sciences, 60 Nanyang Drive, 637551, Singapore. Electronic address: ggrueber@ntu.edu.sg.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Grüber
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008958:Models, Molecular; D010447:Peptide Hydrolases / Q000737:chemistry; D011487:Protein Conformation; D000072417:Protein Domains; D011794:Quercetin / Q000494:pharmacology; D020365:RNA Helicases / Q000737:chemistry; D000077185:Resveratrol; D053838:Scattering, Small Angle; D012697:Serine Endopeptidases / Q000737:chemistry; D012996:Solutions; D013267:Stilbenes / Q000494:pharmacology; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D014961:X-Ray Diffraction; D000071244:Zika Virus / Q000737:chemistry* / Q000201:enzymology; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000975:Antioxidants; D004791:Enzyme Inhibitors; D010447:Peptide Hydrolases; D011794:Quercetin; D020365:RNA Helicases; D000077185:Resveratrol; D012697:Serine Endopeptidases; D012996:Solutions; D013267:Stilbenes; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C582223:NS5 protein, dengue virus
keywords: Flavivirus; Helicase; Methyltransferase; Nonstructural proteins; Protease; Small-angle X-ray scattering; Viral polymerase; Zika