Selective incorporation of vRNP into influenza A virions determined by its specific interaction with M1 protein

Virology. 2017 May:505:23-32. doi: 10.1016/j.virol.2017.02.008. Epub 2017 Feb 17.

Abstract

Influenza A viruses contain eight single-stranded, negative-sense RNA segments as viral genomes in the form of viral ribonucleoproteins (vRNPs). During genome replication in the nucleus, positive-sense complementary RNPs (cRNPs) are produced as replicative intermediates, which are not incorporated into progeny virions. To analyze the mechanism of selective vRNP incorporation into progeny virions, we quantified vRNPs and cRNPs in the nuclear and cytosolic fractions of infected cells, using a strand-specific qRT-PCR. Unexpectedly, we found that cRNPs were also exported to the cytoplasm. This export was chromosome region maintenance 1 (CRM1)-independent unlike that of vRNPs. Although both vRNPs and cRNPs were present in the cytosol, viral matrix (M1) protein, a key regulator for viral assembly, preferentially bound vRNPs over cRNPs. These results indicate that influenza A viruses selectively uptake cytosolic vRNPs through a specific interaction with M1 during viral assembly.

Keywords: Assembly; CRM1; CRNP; Influenza; M1; Nuclear export; Trafficking; VRNP.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • A549 Cells
  • Active Transport, Cell Nucleus
  • Animals
  • Cell Line, Tumor
  • Cell Nucleus / virology
  • Cytoplasm / virology
  • Dogs
  • Exportin 1 Protein
  • Genome, Viral / genetics
  • HEK293 Cells
  • Humans
  • Influenza A virus / genetics*
  • Karyopherins / metabolism
  • Madin Darby Canine Kidney Cells
  • RNA, Viral / genetics*
  • RNA, Viral / metabolism
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ribonucleoproteins / metabolism*
  • Viral Matrix Proteins / genetics
  • Viral Matrix Proteins / metabolism*
  • Virus Assembly / genetics
  • Virus Assembly / physiology*
  • Virus Replication / genetics
  • Virus Replication / physiology*

Substances

  • Karyopherins
  • M1 protein, Influenza A virus
  • RNA, Viral
  • Receptors, Cytoplasmic and Nuclear
  • Ribonucleoproteins
  • Viral Matrix Proteins