Influenza A virus NS2 protein mediates vRNP nuclear export through NES-independent interaction with hCRM1.
For nuclear export of proteins, the formation of a ternary export complex composed of the export substrate, a cellular export factor and Ran-GTP is crucial. CRM1 is a cellular export factor for proteins containing leucine-rich nuclear export signals (NESs). Although the NES sequence is crucial for nuclear export, its exact role in the formation of the ternary export complex is controversial. Here we demonstrate an interaction between human CRM1 (hCRM1) and influenza A virus NS2 protein, which contains an NES motif in its N-terminal region. Replacement of the hydrophobic amino acids in the NES motif did not abolish NS2's interaction with hCRM1. Using our recently established systems for the generation of influenza virus or virus-like particles from cloned cDNAs, we found that NS2 is essential for nuclear export of influenza virus ribonucleoprotein (RNP) complexes, and that alteration of the NS2-NES abrogated this event and influenza virus generation. These findings suggest that the NS2-NES is not crucial for the interaction of this protein with hCRM1, but is for the formation of the ternary export complex with Ran-GTP.
Neumann
G
G
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
Hughes
M T
MT
Kawaoka
Y
Y
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism* / Q000821:virology; D016384:Consensus Sequence; D006801:Humans; D009980:Influenza A virus / Q000502:physiology*; D028884:Karyopherins; D007668:Kidney; D007930:Leucine; D008018:Life Cycle Stages; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000502:physiology; D012318:RNA Polymerase I / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology*; D000097606:Exportin 1 Protein
D002352:Carrier Proteins; D028884:Karyopherins; D007930:Leucine; D009687:Nuclear Proteins; D012318:RNA Polymerase I; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; D000097606:Exportin 1 Protein