Surface expression of influenza virus neuraminidase, an amino-terminally anchored viral membrane glycoprotein, in polarized epithelial cells.
We have investigated the site of surface expression of the neuraminidase (NA) glycoprotein of influenza A virus, which, in contrast to the hemagglutinin, is bound to membranes by hydrophobic residues near the NH2-terminus. Madin-Darby canine kidney or primary African green monkey kidney cells infected with influenza A/WSN/33 virus and subsequently labeled with monoclonal antibody to the NA and then with a colloidal gold- or ferritin-conjugated second antibody exhibited specific labeling of apical surfaces. Using simian virus 40 late expression vectors, we also studied the surface expression of the complete NA gene (SNC) and a truncated NA gene (SN10) in either primary or a polarized continuous line (MA104) of African green monkey kidney cells. The polypeptides encoded by the cloned NA cDNAs were expressed on the surface of both cell types. Analysis of [3H]mannose-labeled polypeptides from recombinant virus-infected MA104 cells showed that the products of cloned NA cDNA comigrated with glycosylated NA from influenza virus-infected cells. Both the complete and the truncated glycoproteins were found to be preferentially expressed on apical plasma membranes, as detected by immunogold labeling. These results indicate that the NA polypeptide contains structural features capable of directing the transport of the protein to apical cell surfaces and the first 10 amino-terminal residues of the NA polypeptide are not involved in this process.
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004247:DNA; D004285:Dogs; D004848:Epithelium / Q000032:analysis / Q000648:ultrastructure*; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000032:analysis; D009980:Influenza A virus / Q000032:analysis* / Q000235:genetics; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D009439:Neuraminidase / Q000032:analysis* / Q000235:genetics; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000032:analysis; D013539:Simian virus 40 / Q000235:genetics; D014721:Vesicular stomatitis Indiana virus / Q000032:analysis; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000235:genetics
D006023:Glycoproteins; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D008565:Membrane Proteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D004247:DNA; D009439:Neuraminidase