title: Canine distemper virus infects canine keratinocytes and immune cells by using overlapping and distinct regions located on one side of the attachment protein.
abstract: The morbilliviruses measles virus (MeV) and canine distemper virus (CDV) both rely on two surface glycoproteins, the attachment (H) and fusion proteins, to promote fusion activity for viral cell entry. Growing evidence suggests that morbilliviruses infect multiple cell types by binding to distinct host cell surface receptors. Currently, the only known in vivo receptor used by morbilliviruses is CD150/SLAM, a molecule expressed in certain immune cells. Here we investigated the usage of multiple receptors by the highly virulent and demyelinating CDV strain A75/17. We based our study on the assumption that CDV-H may interact with receptors similar to those for MeV, and we conducted systematic alanine-scanning mutagenesis on CDV-H throughout one side of the β-propeller documented in MeV-H to contain multiple receptor-binding sites. Functional and biochemical assays performed with SLAM-expressing cells and primary canine epithelial keratinocytes identified 11 residues mutation of which selectively abrogated fusion in keratinocytes. Among these, four were identical to amino acids identified in MeV-H as residues contacting a putative receptor expressed in polarized epithelial cells. Strikingly, when mapped on a CDV-H structural model, all residues clustered in or around a recessed groove located on one side of CDV-H. In contrast, reported CDV-H mutants with SLAM-dependent fusion deficiencies were characterized by additional impairments to the promotion of fusion in keratinocytes. Furthermore, upon transfer of residues that selectively impaired fusion induction in keratinocytes into the CDV-H of the vaccine strain, fusion remained largely unaltered. Taken together, our results suggest that a restricted region on one side of CDV-H contains distinct and overlapping sites that control functional interaction with multiple receptors.
authors.affiliation: Department of Clinical Resarch and Veterinary Public Health, University of Bern, Bremgartenstrasse 109a, 3001 Bern, Switzerland.
authors.forename: Johannes P M
authors.initials: JP
authors.lastname: Langedijk
authors.forename: Jozef
authors.initials: J
authors.lastname: Janda
authors.forename: Francesco C
authors.initials: FC
authors.lastname: Origgi
authors.forename: Claes
authors.initials: C
authors.lastname: Örvell
authors.forename: Marc
authors.initials: M
authors.lastname: Vandevelde
authors.forename: Andreas
authors.initials: A
authors.lastname: Zurbriggen
authors.forename: Philippe
authors.initials: P
authors.lastname: Plattet
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002460:Cell Line; D004217:Distemper Virus, Canine / Q000737:chemistry / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D015603:Keratinocytes / Q000821:virology*; D007962:Leukocytes / Q000821:virology*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011487:Protein Conformation; D011991:Receptors, Virus / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053585:Virus Attachment
chemical_list: D011991:Receptors, Virus; D014764:Viral Proteins