Identification of two amino acids in the hemagglutinin glycoprotein of measles virus (MV) that govern hemadsorption, HeLa cell fusion, and CD46 downregulation: phenotypic markers that differentiate vaccine and wild-type MV strains.
We have used site-directed mutagenesis of the hemagglutinin (H) glycoprotein of measles virus (MV) to investigate the molecular basis for the phenotypic differences observed between MV vaccine strains and recently isolated wild-type MV strains. The former downregulate CD46, the putative cellular receptor of MV, are positive for hemadsorption, and are fusogenic in HeLa cells, whereas the latter are negative for these phenotypic markers. CD46 downregulation in particular, could have profound consequences for the immunopathology of MV infection, as this molecule protects the cell from complement lysis. Mutagenesis of two amino acids, valine and tyrosine at positions 451 and 481, respectively, in the H protein from the vaccine-like Hallé MV strain to their counterparts, glutamate and asparagine, in the H protein from the wild-type Ma93F MV strain (creating the V451E/Y481N double mutation) abrogated CD46 downregulation, HeLa cell fusion, and hemadsorption. The converse double mutagenesis of the Ma93F H protein (E451V/N481Y) transferred the CD46-downregulating, fusogenic, and hemadsorption functions to this protein. The data provide the first mapping study of the functional domains of MV H. The consequences of these results for MV vaccine design and the role of CD46 in MV infection are discussed.
Lecouturier
V
V
Institut National de la Sante et de la Recherche Medicale U.404, Institut Natiional de la Santéet de la Recherche Médicale U.404, Immunité et Vaccination, Insitut Pasteur de Lyon,France.
Fayolle
J
J
Caballero
M
M
Carabaña
J
J
Celma
M L
ML
Fernandez-Muñoz
R
R
Wild
T F
TF
Buckland
R
R
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D015536:Down-Regulation; D006367:HeLa Cells; D006382:Hemadsorption; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D008458:Measles Vaccine / Q000737:chemistry*; D008459:Measles virus / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D051916:Membrane Cofactor Protein; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000378:metabolism*; D010641:Phenotype; D011817:Rabbits; D011991:Receptors, Virus / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014709:Vero Cells
D015703:Antigens, CD; C497857:CD46 protein, human; D006389:Hemagglutinins, Viral; D008458:Measles Vaccine; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; C032397:hemagglutinin protein G, measles virus