Imaging and characterizing influenza A virus mRNA transport in living cells

Nucleic Acids Res. 2008 Sep;36(15):4913-28. doi: 10.1093/nar/gkn475. Epub 2008 Jul 24.

Abstract

The mechanisms of influenza A virus mRNA intracellular transport are still not clearly understood. Here, we visualized the distribution and transport of influenza A virus mRNA in living cells using molecular beacon (MB) technology. Confocal-FRAP measurements determined that the transport of influenza A virus intronless mRNA, in both nucleus and cytoplasm, was energy dependent, being similar to that of Poly(A)(+) RNA. Drug inhibition studies in living cells revealed that the export of influenza A virus mRNA is independent of the CRM1 pathway, while the function of RNA polymerase II (RNAP-II) may be needed. In addition, viral NS1 protein and cellular TAP protein were found associated with influenza A virus mRNA in the cell nucleus. These findings characterize influenza A virus mRNA transport in living cells and suggest that influenza A virus mRNA may be exported from the nucleus by the cellular TAP/p15 pathway with NS1 protein and RNAP-II participation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Nucleus / chemistry
  • Cell Nucleus / virology
  • Cytoplasm / virology
  • Dactinomycin / pharmacology
  • Dogs
  • Exportin 1 Protein
  • Fluorescence Recovery After Photobleaching
  • Immunoprecipitation
  • Influenza A virus / genetics
  • Influenza A virus / metabolism*
  • Karyopherins / metabolism
  • Microscopy, Confocal
  • Nucleocytoplasmic Transport Proteins / analysis
  • RNA Transport / drug effects
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism*
  • RNA, Viral / analysis
  • RNA, Viral / metabolism*
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Viral Nonstructural Proteins / analysis

Substances

  • INS1 protein, influenza virus
  • Karyopherins
  • Nucleocytoplasmic Transport Proteins
  • RNA, Messenger
  • RNA, Viral
  • Receptors, Cytoplasmic and Nuclear
  • Viral Nonstructural Proteins
  • Dactinomycin