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Journal of Virology, January 2004, p. 302-313, Vol. 78, No. 1
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.1.302-313.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Selectively Receptor-Blind Measles Viruses: Identification of Residues Necessary for SLAM- or CD46-Induced Fusion and Their Localization on a New Hemagglutinin Structural Model

Sompong Vongpunsawad,1 Numan Oezgun,2 Werner Braun,2 and Roberto Cattaneo1*

Molecular Medicine Program, Mayo Clinic, and Virology and Gene Therapy, Mayo Graduate School, Rochester, Minnesota 55095,1 Sealy Center for Structural Biology, University of Texas Medical Branch, Galveston, Texas 775552

Received 22 July 2003/ Accepted 23 September 2003

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 ß-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.

* Corresponding author. Mailing address: Molecular Medicine Program, Mayo Clinic, Guggenheim 18, 200 First St., SW, Rochester, MN 55905. Phone: (507) 284-0181. Fax: (507) 266-2122. E-mail: cattaneo.roberto{at}mayo.edu.

Journal of Virology, January 2004, p. 302-313, Vol. 78, No. 1
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.1.302-313.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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