RESUMEN
We identified a new subgroup of koala retrovirus (KoRV), named KoRV-J, which utilizes thiamine transport protein 1 as a receptor instead of the Pit-1 receptor used by KoRV (KoRV-A). By subgroup-specific PCR, KoRV-J and KoRV-A were detected in 67.5 and 100% of koalas originating from koalas from northern Australia, respectively. Altogether, our results indicate that the invasion of the koala population by KoRV-J may have occurred more recently than invasion by KoRV-A.
Asunto(s)
Animales de Zoológico/virología , Retrovirus Endógenos/clasificación , Retrovirus Endógenos/genética , Phascolarctidae/virología , Secuencia de Aminoácidos , Animales , Australia , Secuencia de Bases , ADN Viral/genética , Retrovirus Endógenos/aislamiento & purificación , Evolución Molecular , Genes env , Células HEK293 , Especificidad del Huésped , Humanos , Japón , Datos de Secuencia Molecular , Filogenia , Homología de Secuencia de AminoácidoRESUMEN
RD-114 virus is a replication-competent feline endogenous retrovirus (ERV). RD-114 virus had been thought to be xenotropic; however, recent findings indicate that RD-114 virus is polytropic and can infect and grow efficiently in feline cells. Receptor(s) for RD-114 virus has not been identified and characterized in cats. In this study, we confirmed that two feline sodium-dependent neutral amino acid transporters (ASCTs), fASCT1 and fASCT2, function as RD-114 virus receptors. By chimeric analyses of feline and murine ASCTs, we revealed that extracellular loop 2 of both fASCT1 and fASCT2 determines the susceptibility to RD-114 virus. Further, we revealed ubiquitous expression of these genes, consistent with the general metabolic role of the ASCT molecules. Our study indicates that RD-114 virus may reinfect tissues and cells in cats, once the virus is activated. Implications of the involvement of RD-114 virus in feline oncogenesis are also discussed.
Asunto(s)
Sistema de Transporte de Aminoácidos ASC/metabolismo , Retrovirus Endógenos/patogenicidad , Receptores Virales/metabolismo , Infecciones por Retroviridae/veterinaria , Sistema de Transporte de Aminoácidos ASC/genética , Animales , Enfermedades de los Gatos/virología , Gatos/virología , Línea Celular , Retrovirus Endógenos/metabolismo , Retrovirus Endógenos/fisiología , Ratones , Datos de Secuencia Molecular , Especificidad de Órganos , Receptores Virales/genética , Infecciones por Retroviridae/virología , Análisis de Secuencia de ADN , Activación ViralRESUMEN
Koala retrovirus (KoRV) is a unique gammaretrovirus that is currently endogenizing into its host and considered to be associated with leukemia, lymphoma and immunosuppression in koalas (Phascolactos cinereus). In this study, it was demonstrated that WWP2 or WWP2-like E3 ubiquitin ligases possessing the WW domain closely related to WWP2 and Vps4A/B are involved in KoRV budding. These data suggest that KoRV Gag recruits the cellular endosomal sorting complex required for transport machinery through interaction of the PPPY L-domain with the WW domain(s) of WWP2 and that progeny virions are released from cells by utilizing the multivesicular body sorting pathway.
Asunto(s)
Gammaretrovirus/fisiología , Productos del Gen gag/metabolismo , Interacciones Huésped-Patógeno , Ubiquitina-Proteína Ligasas/metabolismo , Liberación del Virus , Animales , Línea Celular , Humanos , Phascolarctidae , Unión ProteicaRESUMEN
Human skin fibroblasts were cultured in the presence of 4-methylumbelliferone, an inhibitor of hyaluronan synthesis. Gelatinolytic activity excreted in the medium was examined by zymography and gelatinase assay using a fluorogenic substrate. 4-Methylumbelliferone added to the medium activated the latent form of matrix metalloproteinase-2 in a dose- and time-dependent manner. Immunoblot analysis also showed the conversion of the latent form of matrix metalloproteinase-2 to its active form. This activation was observed even when the cells were cultured with both 4-methylumbelliferone and hyaluronan. Addition of Streptomyces hyaluronidase to the medium during cultivation did not activate the latent form of matrix metalloproteinase-2. Reverse transcription-polymerase chain reaction revealed that 4-methylumbelliferone markedly increased the level of mRNA for membrane type 1-matrix metalloproteinase, whereas levels of mRNA for matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 were little affected. These results suggest that 4-methylumbelliferone induces the expression of membrane type 1-matrix metalloproteinase, resulting in activation of matrix metalloproteinase-2, in cultured human skin fibroblasts.