Influenza virus entry and infection require host cell N-linked glycoprotein.
A widely held view of influenza virus infection is that the viral receptor consists of cell surface carbohydrate sialic acid, which can be present as glycoprotein or glycolipid. Here, we examined influenza virus entry and infection in Lec1 cells, a mutant CHO cell line deficient in terminal N-linked glycosylation caused by a mutation in the N-acetylglucosaminyltransferase I (GnT1) gene. We show that influenza virus cannot infect Lec1 cells, despite having full capacity to undergo virus binding and fusion. Lec1 cells also show no virus replication defect, and infection was restored in Lec1 cells expressing wild-type GnT1. Viruses were apparently arrested at the level of internalization from the plasma membrane and were not endocytosed. Lec1 cells were refractory to infection by several strains of influenza virus, including H1 and H3 strains of influenza A, as well as influenza B virus. Finally, cleavage of N-glycans from wild-type CHO cells markedly reduced infection by influenza virus. We suggest that influenza virus specifically requires N-linked glycoprotein for entry into cells, and that sialic acid, although acting as an efficient attachment factor, is not sufficient as an influenza virus receptor in vivo.
Chu
Victor C
VC
Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Whittaker
Gary R
GR
D000818:Animals; D001710:Biotin / Q000737:chemistry; D019921:Biotinylation; D016466:CHO Cells; D002241:Carbohydrates / Q000737:chemistry; D002460:Cell Line; D002478:Cells, Cultured / Q000821:virology; D006224:Cricetinae; D004705:Endocytosis; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006023:Glycoproteins / Q000737:chemistry / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D007251:Influenza, Human / Q000473:pathology*; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009975:Orthomyxoviridae / Q000235:genetics / Q000472:pathogenicity*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000378:metabolism
D002241:Carbohydrates; D006023:Glycoproteins; D014760:Viral Fusion Proteins; D001710:Biotin; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D019158:N-Acetylneuraminic Acid