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Affinity Purification of Influenza Virus Ribonucleoprotein Complexes from the Chromatin of Infected Cells
11:20

Affinity Purification of Influenza Virus Ribonucleoprotein Complexes from the Chromatin of Infected Cells

Published on: June 3, 2012

Oligomerization of the influenza virus polymerase complex in vivo

Núria Jorba1, Estela Area1, Juan Ortín1

  • 1Centro Nacional de Biotecnología (CSIC), Darwin 3, Campus de Cantoblanco, 28049 Madrid, Spain.

The Journal of General Virology
|January 17, 2008
PubMed

Insights

The influenza virus polymerase, essential for virus replication, was purified and found to form higher-order oligomers, not just trimers. This finding suggests new insights into influenza virus infection mechanisms.

Area of Science:

  • Virology
  • Molecular Biology
  • Protein Biochemistry

Background:

  • The influenza virus polymerase complex, a trimer of PB1, PB2, and PA subunits, is crucial for viral transcription and replication.
  • Understanding the native structure and assembly of this complex is key to developing antiviral strategies.

Purpose of the Study:

  • To express and purify the functional influenza virus polymerase complex from human cells.
  • To investigate the oligomeric state of the intracellular influenza virus polymerase complex.

Main Methods:

  • Co-transfection of influenza virus polymerase subunit cDNAs into human cells.
  • Tandem Affinity Purification (TAP) using IgG-Sepharose and calmodulin-agarose chromatography.
  • Gel-filtration analysis to determine the molecular size and oligomeric state of the purified complex.

Main Results:

  • The influenza virus polymerase complex was successfully purified from human cells.
  • Gel-filtration analysis revealed that while most polymerase existed as a heterotrimer, a significant portion formed dimers, trimers, and higher-order oligomers.
  • Experiments with alternatively tagged subunits confirmed the intracellular formation of polymerase oligomers.

Conclusions:

  • The influenza virus polymerase complex can exist as higher-order oligomers within infected cells.
  • These oligomeric forms may have implications for viral transcription, replication, and overall virus infection dynamics.
  • Further research is warranted to elucidate the functional significance of these oligomers in the viral life cycle.

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