RESUMEN
We analyzed the roles of the individual measles virus (MV) surface glycoproteins in mediating functional and structural interactions with human CD46, the primary MV receptor. On one cell population, recombinant vaccinia virus vectors were used to produce the MV hemagglutinin (H) and fusion (F) glycoproteins. As fusion partner cells, various cell types were examined, without or with human CD46 (endogenous or recombinant vaccinia virus encoded). Fusion between the two cell populations was monitored by a quantitative reporter gene activation assay and by syncytium formation. MV glycoproteins promoted fusion with primate cells but not with nonprimate cells; recombinant CD46 rendered nonprimate cells competent for MV glycoprotein-mediated fusion. Markedly different fusion specificity was observed for another morbillivirus, canine distemper virus (CDV): recombinant CDV glycoproteins promoted fusion with primate and nonprimate cells independently of CD46. Fusion by the recombinant MV and CDV glycoproteins required coexpression of H plus F in either homologous or heterologous combinations. To assess the role of H versus F in determining the CD46 dependence of MV fusion, we examined the fusion specificities of cells producing heterologous glycoprotein combinations. The specificity of HMV plus FCDV paralleled that observed for the homologous MV glycoproteins: fusion occurred with primate cells but not with nonprimate cells unless they produced recombinant CD46. By contrast, the specificity of HCDV plus FMV paralleled that for the homologous CDV glycoproteins: fusion occurred with either primate or nonprimate cells with no dependence on CD46. Thus, for both MV and CDV, fusion specificity was determined by H. In particular, the results demonstrate a functional interaction between HMV and CD46. Flow cytometry and antibody coprecipitation studies provided a structural correlate to this functional interaction: CD46 formed a molecular complex with HMV but not with FMV or with either CDV glycoprotein. These results highlight the critical role of the H glycoprotein in determining MV specificity for CD46-positive cells.
Asunto(s)
Antígenos CD/metabolismo , Hemaglutininas Virales/metabolismo , Virus del Sarampión/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptores Virales/metabolismo , Células 3T3 , Animales , Secuencia de Bases , Fusión Celular , Cartilla de ADN , Virus del Moquillo Canino/metabolismo , Glicoproteínas/metabolismo , Células HeLa , Hemaglutininas Virales/genética , Humanos , Virus del Sarampión/genética , Proteína Cofactora de Membrana , Ratones , Datos de Secuencia Molecular , Unión Proteica , Relación Estructura-Actividad , Virus Vaccinia/genéticaRESUMEN
Measles virus (MV) and canine distemper virus (CDV) are morbilliviruses that cause acute illnesses and several persistent central nervous system infections in humans and in dogs, respectively. Characteristically, the cytopathic effect of these viruses is the formation of syncytia in permissive cells. In this study, a vaccinia virus expression system was used to express MV and CDV hemagglutinin (HA) and fusion (F) envelope proteins. We found that cotransfecting F and HA genes of MV or F and HA genes of CDV resulted in extensive syncytium formation in permissive cells while transfecting either F or HA alone did not. Similar experiments with heterologous pairs of proteins, CDV-F with MV-HA or MV-F with CDV-HA, caused significant cell fusion in both cases. These results indicate that in this expression system, cell fusion requires both F and HA; however, the functions of these proteins are interchangeable between the two types of morbilliviruses. Human-mouse somatic hybrids were used to determine the human chromosome conferring susceptibility to either MV and CDV. Of the 12 hybrids screened, none were sensitive to MV. Two of the hybrids containing human chromosome 19 formed syncytia following CDV infection. In addition, these two hybrids underwent cell fusion when cotransfected with CDV-F and CDV-HA (but not MV-F and MV-HA) glycoproteins by using the vaccinia virus expression system. To discover the viral component responsible for cell specificity, complementation experiments coexpressing CDV-HA with MV-F or CDV-F with MV-HA in the CDV-sensitive hybrids were performed. We found that syncytia were formed only in the presence of CDV-HA. These results support the idea that the HA protein is responsible for cell tropism. Furthermore, while the F protein is necessary for the fusion process, it is interchangeable with the F protein from other morbilliviruses.
Asunto(s)
Virus del Moquillo Canino/crecimiento & desarrollo , Hemaglutininas Virales/metabolismo , Virus del Sarampión/crecimiento & desarrollo , Proteínas Virales de Fusión/metabolismo , Animales , Secuencia de Bases , Fusión Celular , Cartilla de ADN/química , Genes Virales , Prueba de Complementación Genética , Hemaglutininas Virales/genética , Humanos , Células Híbridas , Técnicas In Vitro , Ratones , Datos de Secuencia Molecular , Proteínas Virales de Fusión/genética , Proteínas Estructurales Virales/genéticaRESUMEN
The late Ilse Bry made numerous contributions to health science librarianship in the roles of scholar and librarian. Her professional interests lay in the literature and bibliography of the behavioral sciences; she served as Librarian of the New York Psychoanalytic Institute and later became the library associate in charge of the New York University Psychiatric Library. Upon leaving active librarianship Ilse Bry continued to work as a meticulous bibliographer and as Editor of the Mental Health Book Review Index.