Dengue Virus Nonstructural Protein 5 (NS5) Assembles into a Dimer with a Unique Methyltransferase and Polymerase Interface

Valerie J. Klema, Mengyi Ye, Aditya Hindupur, Tadahisa Teramoto, Keerthi Gottipati, Radhakrishnan Padmanabhan, Kyung Choi

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Flavivirus nonstructural protein 5 (NS5) consists of methyltransferase (MTase) and RNA-dependent RNA polymerase (RdRp) domains, which catalyze 5’-RNA capping/methylation and RNA synthesis, respectively, during viral genome replication. Although the crystal structure of flavivirus NS5 is known, no data about the quaternary organization of the functional enzyme are available. We report the crystal structure of dengue virus full-length NS5, where eight molecules of NS5 are arranged as four independent dimers in the crystallographic asymmetric unit. The relative orientation of each monomer within the dimer, as well as the orientations of the MTase and RdRp domains within each monomer, is conserved, suggesting that these structural arrangements represent the biologically relevant conformation and assembly of this multi-functional enzyme. Essential interactions between MTase and RdRp domains are maintained in the NS5 dimer via inter-molecular interactions, providing evidence that flavivirus NS5 can adopt multiple conformations while preserving necessary interactions between the MTase and RdRp domains. Furthermore, many NS5 residues that reduce viral replication are located at either the inter-domain interface within a monomer or at the inter-molecular interface within the dimer. Hence the X-ray structure of NS5 presented here suggests that MTase and RdRp activities could be coordinated as a dimer during viral genome replication.

Original languageEnglish (US)
Article numbere1005451
JournalPLoS Pathogens
Volume12
Issue number2
DOIs
StatePublished - Feb 1 2016

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Dengue Virus
Methyltransferases
RNA Replicase
Flavivirus
Proteins
Viral Genome
RNA
Enzymes
Methylation
X-Rays

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Dengue Virus Nonstructural Protein 5 (NS5) Assembles into a Dimer with a Unique Methyltransferase and Polymerase Interface. / Klema, Valerie J.; Ye, Mengyi; Hindupur, Aditya; Teramoto, Tadahisa; Gottipati, Keerthi; Padmanabhan, Radhakrishnan; Choi, Kyung.

In: PLoS Pathogens, Vol. 12, No. 2, e1005451, 01.02.2016.

Research output: Contribution to journalArticle

Klema, Valerie J. ; Ye, Mengyi ; Hindupur, Aditya ; Teramoto, Tadahisa ; Gottipati, Keerthi ; Padmanabhan, Radhakrishnan ; Choi, Kyung. / Dengue Virus Nonstructural Protein 5 (NS5) Assembles into a Dimer with a Unique Methyltransferase and Polymerase Interface. In: PLoS Pathogens. 2016 ; Vol. 12, No. 2.
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