title: The receptor attachment function of measles virus hemagglutinin can be replaced with an autonomous protein that binds Her2/neu while maintaining its fusion-helper function.
abstract: Cell entry of enveloped viruses is initiated by attachment to the virus receptor followed by fusion between the virus and host cell membranes. Measles virus (MV) attachment to its receptor is mediated by the hemagglutinin (H), which is thought to produce conformational changes in the membrane fusion protein (F) that trigger insertion of its fusion peptide into the target cell membrane. Here, we uncoupled receptor attachment and the fusion-helper function of H by introducing Y481A, R533A, S548L, and F549S mutations into the viral attachment protein that made it blind to its normal receptors. An artificial receptor attachment protein specific for Her2/neu was incorporated into the membranes of pseudotyped lentivirus particles as a separate transmembrane protein along with the F protein. Surprisingly, these particles entered efficiently into Her2/neu-positive SK-OV-3 as well as CHO-Her2 cells. Cell entry was independent of endocytosis but strictly dependent on the presence of H. H-specific monoclonal antibodies, as well as a mutation in H interfering with H/F cooperation, blocked cell entry. The particles mediated stable and specific transfer of reporter genes into Her2/neu-positive human tumor cells also in vivo, while exhibiting improved infectivity and higher titers than Her2/neu-targeted vectors displaying the targeting domain on H. Extending the current model of MV cell entry, the data suggest that receptor binding of H is not required for its fusion-helper function but that particle-cell contact in general may be sufficient to induce the conformational changes in the H/F complex and activate membrane fusion.
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Rasbach
authors.forename: Tobias
authors.initials: T
authors.lastname: Abel
authors.forename: Robert C
authors.initials: RC
authors.lastname: Münch
authors.forename: Klaus
authors.initials: K
authors.lastname: Boller
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schneider-Schaulies
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Buchholz
mesh_terms: D000818:Animals; D002460:Cell Line; D005260:Female; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008457:Measles / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D008459:Measles virus / Q000235:genetics / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D053585:Virus Attachment; D053586:Virus Internalization
chemical_list: D006389:Hemagglutinins, Viral; D018719:Erb-b2 Receptor Tyrosine Kinases; D011991:Receptors, Virus
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