title: Single amino acid changes in the Nipah and Hendra virus attachment glycoproteins distinguish ephrinB2 from ephrinB3 usage.
abstract: The henipaviruses, Nipah virus (NiV) and Hendra virus (HeV), are lethal emerging paramyxoviruses. EphrinB2 and ephrinB3 have been identified as receptors for henipavirus entry. NiV and HeV share similar cellular tropisms and likely use an identical receptor set, although a quantitative comparison of receptor usage by NiV and HeV has not been reported. Here we show that (i) soluble NiV attachment protein G (sNiV-G) bound to cell surface-expressed ephrinB3 with a 30-fold higher affinity than that of sHeV-G, (ii) NiV envelope pseudotyped reporter virus (NiVpp) entered ephrinB3-expressing cells much more efficiently than did HeV pseudotyped particles (HeVpp), and (iii) NiVpp but not HeVpp entry was inhibited efficiently by soluble ephrinB3. These data underscore the finding that NiV uses ephrinB3 more efficiently than does HeV. Henipavirus G chimeric protein analysis implicated residue 507 in the G ectodomain in efficient ephrinB3 usage. Curiously, alternative versions of published HeV-G sequences show variations at residue 507 that can clearly affect ephrinB3 but not ephrinB2 usage. We further defined surrounding mutations (W504A and E505A) that diminished ephrinB3-dependent binding and viral entry without compromising ephrinB2 receptor usage and another mutation (E533Q) that abrogated both ephrinB2 and -B3 usage. Our results suggest that ephrinB2 and -B3 binding determinants on henipavirus G are distinct and dissociable. Global expression analysis showed that ephrinB3, but not ephrinB2, is expressed in the brain stem. Thus, ephrinB3-mediated viral entry and pathology may underlie the severe brain stem neuronal dysfunction seen in fatal Nipah viral encephalitis. Characterizing the determinants of ephrinB2 versus -B3 usage will further our understanding of henipavirus pathogenesis.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, UCLA AIDS Institute, 609 Charles Young Dr., 3825 Molecular Science Building, Los Angeles, CA 90095, USA.
authors.forename: Oscar A
authors.initials: OA
authors.lastname: Negrete
authors.forename: David
authors.initials: D
authors.lastname: Chu
authors.forename: Hector C
authors.initials: HC
authors.lastname: Aguilar
authors.forename: Benhur
authors.initials: B
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D036388:Ephrin-B2 / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D036389:Ephrin-B3 / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D045404:Hendra Virus / Q000502:physiology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D045405:Nipah Virus / Q000502:physiology*; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D012694:Serine / Q000737:chemistry / Q000235:genetics; D013912:Threonine / Q000737:chemistry / Q000235:genetics; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D036388:Ephrin-B2; D036389:Ephrin-B3; D014759:Viral Envelope Proteins; C116204:attachment protein G; D013912:Threonine; D012694:Serine