Expression of measles virus nucleoprotein induces apoptosis and modulates diverse functional proteins in cultured mammalian cells

PLoS One. 2011 Apr 14;6(4):e18765. doi: 10.1371/journal.pone.0018765.

Abstract

Background: Measles virus nucleoprotein (N) encapsidates the viral RNA, protects it from endonucleases and forms a virus specific template for transcription and replication. It is the most abundant protein during viral infection. Its C-terminal domain is intrinsically disordered imparting it the flexibility to interact with several cellular and viral partners.

Principal findings: In this study, we demonstrate that expression of N within mammalian cells resulted in morphological transitions, nuclear condensation, DNA fragmentation and activation of Caspase 3 eventuating into apoptosis. The rapid generation of intracellular reactive oxygen species (ROS) was involved in the mechanism of cell death. Addition of ascorbic acid (AA) or inhibitor of caspase-3 in the extracellular medium partially reversed N induced apoptosis. We also studied the protein profile of cells expressing N protein. MS analysis revealed the differential expression of 25 proteins out of which 11 proteins were up regulated while 14 show signs of down regulation upon N expression. 2DE results were validated by real time and semi quantitative RT-PCR analysis.

Conclusion: These results show the pro-apoptotic effects of N indicating its possible development as an apoptogenic tool. Our 2DE results present prima facie evidence that the MV nucleoprotein interacts with or causes differential expression of a wide range of cellular factors. At this stage it is not clear as to what the adaptive response of the host cell is and what reflects a strategic modulation exerted by the virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / physiology*
  • Caspase 3 / metabolism
  • Cell Line, Tumor
  • Electrophoresis, Gel, Two-Dimensional
  • Enzyme Activation
  • Humans
  • Mass Spectrometry
  • Measles virus / metabolism*
  • Nucleoproteins / metabolism*
  • Nucleoproteins / physiology
  • Reactive Oxygen Species / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Viral Proteins / metabolism*
  • Viral Proteins / physiology

Substances

  • Nucleoproteins
  • Reactive Oxygen Species
  • Viral Proteins
  • Caspase 3