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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
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Affinity Purification of Influenza Virus Ribonucleoprotein Complexes from the Chromatin of Infected Cells
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Published on: June 3, 2012

A functional sequence-specific interaction between influenza A virus genomic RNA segments.

Cyrille Gavazzi1, Matthieu Yver, Catherine Isel

  • 1Architecture et Réactivité de l'ARN, Université de Strasbourg, Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire, 67084 Strasbourg, France.

Proceedings of the National Academy of Sciences of the United States of America
|September 27, 2013
PubMed
Summary

Scientists discovered a specific RNA interaction in avian influenza A virus that aids in packaging viral genomes. This interaction is crucial for optimal viral replication and influences the composition of viral particles.

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Published on: November 1, 2011

Area of Science:

  • Virology
  • Molecular Biology
  • Genomics

Background:

  • Influenza A viruses cause significant global health issues annually and periodically lead to severe pandemics.
  • The segmented genome of influenza A viruses, while advantageous for evolution, presents challenges in packaging viral RNA (vRNA) into new virions.
  • A selective packaging mechanism is hypothesized but its molecular underpinnings are not fully understood.

Purpose of the Study:

  • To identify and characterize molecular interactions between viral genomic RNA segments involved in selective packaging.
  • To determine the role of identified RNA-RNA interactions in viral replication and particle assembly.
  • To investigate the conservation and implications of these interactions for influenza A virus evolution.

Main Methods:

  • In vitro biochemical assays to detect direct intermolecular RNA-RNA interactions.
  • Construction and analysis of silent trans-complementary mutants in infected cells.
  • Assessment of viral particle production, viral RNA content, and hemagglutinin (HA) titer.
  • Competition experiments to evaluate the effect on copackaging of interacting RNA segments.

Main Results:

  • A direct intermolecular interaction between two specific avian influenza A virus genomic RNA segments was identified.
  • This interaction was confirmed to occur within infected cells and is essential for optimal viral replication.
  • Disruption of the interaction led to increased empty viral particles and reduced vRNA content per particle, without affecting overall particle production.
  • The identified interaction regions are novel packaging signals and are not broadly conserved across influenza A virus strains.

Conclusions:

  • Viral RNA segments can form a sequence-dependent supramolecular network, facilitating selective genome packaging.
  • The identified RNA-RNA interaction promotes the copackaging of specific vRNA segments.
  • The limited conservation of these interactions may restrict genetic reassortment between different influenza A virus strains.