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Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
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Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins

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Rotavirus prevents the expression of host responses by blocking the nucleocytoplasmic transport of polyadenylated

Rosa M Rubio1, Silvia I Mora, Pedro Romero

  • 1Departamento de Génetica del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, Mexico.

Journal of Virology
|March 29, 2013
PubMed

Insights

Rotaviruses shut down host protein synthesis by trapping viral mRNAs in the nucleus. This nuclear retention of polyadenylated messenger RNAs (mRNAs) hinders host antiviral responses during infection.

Area of Science:

  • Virology
  • Molecular Biology
  • Cellular Biology

Background:

  • Rotaviruses are a major cause of severe gastroenteritis in children.
  • During infection, rotaviruses inhibit host cell protein synthesis while promoting viral protein production.
  • The viral protein NSP3 sequesters cytoplasmic poly(A)-binding protein in the nucleus.

Purpose of the Study:

  • To investigate the mechanism by which rotaviruses control host cell translation.
  • To determine the fate of polyadenylated mRNAs in rotavirus-infected cells.
  • To elucidate the role of nuclear retention in rotavirus-mediated translation shutoff.

Main Methods:

  • Analysis of poly(A)-containing mRNA localization in infected cells.
  • Hyperadenylation assessment of nuclear mRNAs.
  • Reporter gene assays using viral and cellular untranslated regions (UTRs).
  • Cytoplasmic vs. nuclear delivery of reporter mRNAs.

Main Results:

  • Poly(A)-containing mRNAs accumulate and become hyperadenylated in the nuclei of rotavirus-infected cells.
  • Viral UTRs do not dictate mRNA translation efficiency in infected cells.
  • Directly delivered cytoplasmic reporter mRNAs are translated, while plasmid-derived mRNAs are poorly translated.

Conclusions:

  • Nuclear retention of poly(A)-containing mRNAs is a key strategy employed by rotaviruses.
  • This mechanism contributes to the shutoff of host cell translation.
  • Nuclear mRNA retention helps suppress host antiviral and stress responses.

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