title: Structural insight and flexible features of NS5 proteins from all four serotypes of Dengue virus in solution.
abstract: Infection by the four serotypes of Dengue virus (DENV-1 to DENV-4) causes an important arthropod-borne viral disease in humans. The multifunctional DENV nonstructural protein 5 (NS5) is essential for capping and replication of the viral RNA and harbours a methyltransferase (MTase) domain and an RNA-dependent RNA polymerase (RdRp) domain. In this study, insights into the overall structure and flexibility of the entire NS5 of all four Dengue virus serotypes in solution are presented for the first time. The solution models derived revealed an arrangement of the full-length NS5 (NS5FL) proteins with the MTase domain positioned at the top of the RdRP domain. The DENV-1 to DENV-4 NS5 forms are elongated and flexible in solution, with DENV-4 NS5 being more compact relative to NS5 from DENV-1, DENV-2 and DENV-3. Solution studies of the individual MTase and RdRp domains show the compactness of the RdRp domain as well as the contribution of the MTase domain and the ten-residue linker region to the flexibility of the entire NS5. Swapping the ten-residue linker between DENV-4 NS5FL and DENV-3 NS5FL demonstrated its importance in MTase-RdRp communication and in concerted interaction with viral and host proteins, as probed by amide hydrogen/deuterium mass spectrometry. Conformational alterations owing to RNA binding are presented.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore.
authors.forename: Wuan Geok
authors.initials: WG
authors.lastname: Saw
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Tria
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore.
authors.forename: Ardina
authors.initials: A
authors.lastname: Grüber
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore.
authors.forename: Malathy Sony
authors.initials: MS
authors.lastname: Subramanian Manimekalai
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, 8 College Road, Singapore 169857, Singapore.
authors.forename: Yongqian
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Biological Sciences, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore.
authors.forename: Arun
authors.initials: A
authors.lastname: Chandramohan
authors.affiliation: Department of Biological Sciences, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore.
authors.forename: Ganesh
authors.initials: G
authors.lastname: Srinivasan Anand
authors.affiliation: Stanford Synchrotron Radiation Lightsource, Stanford Linear Accelerator Center National Laborator, Menlo Park, California, USA.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Matsui
authors.affiliation: Stanford Synchrotron Radiation Lightsource, Stanford Linear Accelerator Center National Laborator, Menlo Park, California, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Weiss
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, 8 College Road, Singapore 169857, Singapore.
authors.forename: Subhash G
authors.initials: SG
authors.lastname: Vasudevan
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Grüber
mesh_terms: D000595:Amino Acid Sequence; D003715:Dengue / Q000821:virology*; D003716:Dengue Virus / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D053838:Scattering, Small Angle; D016415:Sequence Alignment; D065288:Serogroup; D017361:Viral Nonstructural Proteins / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: C582223:NS5 protein, dengue virus; D017361:Viral Nonstructural Proteins
keywords: dengue; flavivirus; methyltransferase; nonstructural proteins; protein flexibility; small-angle X-ray scattering; viral polymerase