Differential Extractability of Influenza Virus Hemagglutinin during Intracellular Transport in Polarized Epithelial Cells and Nonpolar Fibroblasts

• Published in: The Journal of cell biology Vol. 108; no. 3; pp. 821 - 832
• Main Authors: Skibbens, Jeanne E., Roth, Michael G., Matlin, Karl S.
• Format: Journal Article
• Language: English
• Published: New York, NY Rockefeller University Press 01.03.1989; The Rockefeller University Press
• Subjects: 1-Deoxynojirimycin; Animals; Biological and medical sciences; Biological Transport; Cell Line; Cell lines; Cell Membrane - analysis; Cell membranes; Cell physiology; Cells; Chemical Phenomena; Chemistry; Cultured cells; Disulfides; Epithelial cells; Epithelium; Fibroblasts; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Gels; Glucosamine - analogs & derivatives; Glucosamine - pharmacology; Golgi Apparatus - analysis; hemagglutinins; Hemagglutinins, Viral - analysis; Influenza A virus - physiology; influenza virus; Kidney cells; LLC PK1 cells; Membrane and intracellular transports; Molecular and cellular biology; Orthomyxoviridae; Solubility; Lectin; Membrane protein; Hemagglutinin; Immunocytochemistry; Orthomyxoviridae; Polarity; Aves; Dog; Fissipedia; Intracellular transport; Carnivora; Radiolabelling; Secretion; Solubility; Glycoproteins; Influenzavirus; Virus; Vertebrata; Mammalia; Plasma membrane; Epithelial cell; Chicken; Immunofluorescence; Fibroblast; Fluorescence microscopy
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.108.3.821

Biochemical changes in the influenza virus hemagglutinin during intracellular transport to the apical plasma membrane of epithelial cells were investigated in Madin-Darby canine kidney (MDCK) cells and in LLC-PK1 cells stably transfected with a hemagglutinin gene. After pulse-labeling a substantial fraction of hemagglutinin was observed to become insoluble in isotonic solutions of Triton X-100. Insolubility of hemagglutinin was detected late in the transport pathway after addition of complex sugars in the Golgi complex but before insertion of the protein in the plasma membrane. Insolubility was not dependent on oligosaccharide modification since deoxymannojirimycin (dMM), which inhibits mannose trimming, failed to prevent its onset. Insolubility was not due to assembly of virus particles at the plasma membrane because insoluble hemagglutinin was also observed in transfected cells. Hemagglutinin insolubility was also seen in MDCK cells cultured in suspension and in chick embryo fibroblasts, indicating that insolubility and plasma membrane polarity are not simply correlated. In addition to insolubility, an apparent transport-dependent reduction of the disulfide bond linking HA1 and HA2 in hemagglutinin was detected. Because of the timing of both insolubility and the loss of the disulfide bond, these modifications may be important in the delivery of the hemagglutinin to the cell surface.