Influenza A virus NP protein expressed in insect cells by a recombinant baculovirus is associated with a protein kinase activity and possesses single-stranded RNA binding activity

Virus Res. 1992 Jun;24(1):91-106. doi: 10.1016/0168-1702(92)90033-6.

Abstract

Influenza A virus NP protein, the phosphoprotein associated with viral RNA in ribonucleoprotein (RNP) complexes, has been expressed at high levels (approximately 100 mg/liter cells) in insect (Sf9) cells by a baculovirus recombinant, and was localized almost entirely in the nuclei of these cells. NP was purified by immuno-affinity chromatography, and purified NP was shown to autophosphorylate and to phosphorylate casein in a cAMP-independent reaction. Furthermore, purified NP was able to bind to ssRNA as demonstrated by a mobility shift of ssRNA in non-denaturing gels. The binding of NP to ssRNA caused a diminution of its kinase activity in proportion to binding.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Baculoviridae / metabolism
  • Cell Line
  • Genetic Vectors
  • Influenza A virus / genetics
  • Influenza A virus / metabolism*
  • Moths
  • Nucleocapsid Proteins
  • Nucleoproteins*
  • Phosphorylation
  • Protein Kinases / metabolism*
  • RNA, Viral / metabolism*
  • Viral Core Proteins / biosynthesis
  • Viral Core Proteins / genetics
  • Viral Core Proteins / isolation & purification
  • Viral Core Proteins / metabolism*

Substances

  • Nucleocapsid Proteins
  • Nucleoproteins
  • RNA, Viral
  • Viral Core Proteins
  • Protein Kinases