Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein

Arch Virol. 2011 Dec;156(12):2163-72. doi: 10.1007/s00705-011-1110-0. Epub 2011 Sep 24.

Abstract

Groundnut bud necrosis virus belongs to the genus Tospovirus, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in E. coli and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated in vitro by Nicotiana benthamiana plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family Bunyaviridae. The possible implications of the present findings for the viral life cycle are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arachis / virology*
  • Base Sequence
  • Binding Sites
  • Biophysical Phenomena
  • DNA Primers / genetics
  • Molecular Sequence Data
  • Nucleocapsid Proteins / chemistry
  • Nucleocapsid Proteins / genetics
  • Nucleocapsid Proteins / metabolism*
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Phosphorylation
  • Protein Multimerization
  • RNA, Viral / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Tospovirus / genetics
  • Tospovirus / metabolism*
  • Tospovirus / pathogenicity

Substances

  • DNA Primers
  • Nucleocapsid Proteins
  • Peptide Fragments
  • RNA, Viral
  • Recombinant Proteins
  • nucleocapsid protein, Tospovirus