Selectively receptor-blind measles viruses: Identification of residues necessary for SLAM- or CD46-induced fusion and their localization on a new hemagglutinin structural model.
Measles virus (MV) enters cells either through the signaling lymphocyte activation molecule SLAM (CD150) expressed only in immune cells or through the ubiquitously expressed regulator of complement activation, CD46. To identify residues on the attachment protein hemagglutinin (H) essential for fusion support through either receptor, we devised a strategy based on analysis of morbillivirus H-protein sequences, iterative cycles of mutant protein production followed by receptor-based functional assays, and a novel MV H three-dimensional model. This model uses the Newcastle disease virus hemagglutinin-neuraminidase protein structure as a template. We identified seven amino acids important for SLAM- and nine for CD46 (Vero cell receptor)-induced fusion. The MV H three-dimensional model suggests (i) that SLAM- and CD46-relevant residues are located in contiguous areas in propeller beta-sheets 5 and 4, respectively; (ii) that two clusters of SLAM-relevant residues exist and that they are accessible for receptor contact; and (iii) that several CD46-relevant amino acids may be shielded from direct receptor contacts. It appears likely that certain residues support receptor-specific H-protein conformational changes. To verify the importance of the H residues identified with the cell-cell fusion assays for virus entry into cells, we transferred the relevant mutations into genomic MV cDNAs. Indeed, we were able to recover recombinant viruses, and we showed that these replicate selectively in cells expressing SLAM or CD46. Selectively receptor-blind viruses will be used to study MV pathogenesis and may have applications for the production of novel vaccines and therapeutics.
Vongpunsawad
Sompong
S
Molecular Medicine Program, Mayo Clinic, and Virology and Gene Therapy, Mayo Graduate School, Rochester, Minnesota 55095, USA.
Oezgun
Numan
N
Braun
Werner
W
Cattaneo
Roberto
R
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D016466:CHO Cells; D002459:Cell Fusion; D002522:Chlorocebus aethiops; D006224:Cricetinae; D006023:Glycoproteins / Q000378:metabolism*; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism*; D008459:Measles virus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D051916:Membrane Cofactor Protein; D008561:Membrane Fusion / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011956:Receptors, Cell Surface; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1; D014709:Vero Cells
D015703:Antigens, CD; C497857:CD46 protein, human; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D007136:Immunoglobulins; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C000604059:SLAMF1 protein, human; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1