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Parainfluenza virus 5 m protein interaction with host protein 14-3-3 negatively affects virus particle formation

Zifei Pei1, Megan S Harrison, Anthony P Schmitt

  • 1Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802, USA.

Journal of Virology
|December 15, 2010
PubMed

Insights

Parainfluenza virus 5 M protein binds to 14-3-3 protein via phosphorylation at S369, which impairs virus budding. Disrupting this interaction enhances virus assembly and release.

Area of Science:

  • Virology
  • Molecular Biology
  • Cell Biology

Background:

  • Paramyxovirus matrix (M) proteins are crucial for viral assembly, mediating interactions between viral components and cellular membranes.
  • Understanding M protein function is key to controlling viral replication and assembly.

Purpose of the Study:

  • To identify binding partners of the parainfluenza virus 5 (PIV5) M protein.
  • To elucidate the role of M protein interactions in PIV5 assembly and budding.

Main Methods:

  • Yeast two-hybrid screening to identify M protein interactors.
  • Coimmunoprecipitation to confirm protein-protein interactions in vitro and in infected cells.
  • Site-directed mutagenesis to investigate the role of specific M protein residues.
  • Radiolabeling to detect protein phosphorylation.
  • Analysis of virus-like particle (VLP) and recombinant virus budding.

Main Results:

  • 14-3-3 protein was identified as a PIV5 M protein binding partner.
  • Binding occurs through a phosphorylated serine residue (S369) in the M protein's C-terminus.
  • Mutation of S369 to alanine abolished 14-3-3 binding and significantly enhanced VLP and virus budding.
  • Overexpression of 14-3-3 protein inhibited VLP budding.
  • Phosphorylation of M protein at S369 was detected in infected cells and was absent in S369A mutants.
  • Disruption of 14-3-3 binding promoted M protein assembly at the cell surface.

Conclusions:

  • PIV5 M protein is phosphorylated at S369, enabling binding to 14-3-3 proteins.
  • This interaction sequesters M protein away from assembly sites, thereby inhibiting virus budding.
  • Targeting the M protein-14-3-3 interaction could be a strategy to control PIV5 spread.

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