Nuclear MxA Proteins Form a Complex With Influenza Virus NP and Inhibit the Transcription of the Engineered Influenza Virus Genome

Nucleic Acids Res. 2004 Jan 29;32(2):643-52. doi: 10.1093/nar/gkh192. Print 2004.

Abstract

Mx proteins belong to the dynamin superfamily of high molecular weight GTPases and interfere with multiplication of a wide variety of viruses. Earlier studies show that nuclear mouse Mx1 and human MxA designed to be localized in the nucleus inhibit the transcription step of the influenza virus genome. Here we set a transient influenza virus transcription system using luciferase as a reporter gene and cells expressing the three RNA polymerase subunits, PB1, PB2 and PA, and NP. We used this reporter assay system and nuclear-localized MxA proteins to get clues for elucidating the anti-influenza virus activity of MxA. Nuclear-localized VP16-MxA and MxA-TAg NLS strongly interfered with the influenza virus transcription. Over-expression of PB2 led to a slight resumption of the transcription inhibition by nuclear MxA, whereas over-expression of PB1 and PA did not affect the MxA activity. Of interest is that the inhibitory activity of the nuclear MxA was markedly neutralized by over-expression of NP. An NP devoid of its C-terminal region, but containing the N-terminal RNA binding domain, also neutralized the VP16-MxA activity in a dose-dependent manner, whereas an NP lacking the N-terminal region did not affect the VP16-MxA activity. Further, not only VP16-MxA but also the wild-type MxA was found to interact with NP in influenza virus-infected cells. This indicates that the nuclear MxA suppresses the influenza virus transcription by interacting with not only PB2 but also NP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus / metabolism*
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism
  • GTP-Binding Proteins / genetics
  • GTP-Binding Proteins / metabolism*
  • Gene Expression Regulation, Viral
  • Genes, Reporter / genetics
  • Genetic Engineering*
  • Genome, Viral*
  • Humans
  • Mice
  • Myxovirus Resistance Proteins
  • Nucleoproteins / genetics
  • Nucleoproteins / metabolism*
  • Orthomyxoviridae / genetics*
  • Protein Binding
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • RNA, Viral / biosynthesis
  • RNA, Viral / genetics
  • Swiss 3T3 Cells
  • Transcription, Genetic*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • MX1 protein, human
  • Mx1 protein, mouse
  • Myxovirus Resistance Proteins
  • Nucleoproteins
  • Protein Subunits
  • RNA, Viral
  • Viral Proteins
  • human parainfluenza virus type 1 nucleoprotein
  • DNA-Directed RNA Polymerases
  • GTP-Binding Proteins