Proceedings of the National Academy of Sciences · 1994 · 593 citations · 42 references
Life SciencesBiochemistryMolecular BiologyGene ExpressionMolecular VirologyVirologyAnimal VirusesCell-surface ReceptorsIon PermeationMembrane ReceptorsMembrane-spanning DomainsPhosphate ExchangePhosphate Transport ProteinsSodium Transport ProteinsSodium-phosphate CotransportTransporter Gene Expression RegulationViral Envelope ProteinsVoltage-clamp Analysis
Cell surface receptors for gibbon ape leukemia virus (Glvr-1) and murine amphotropic retrovirus (Ram-1) are distinct but related proteins having multiple membrane-spanning regions. Distant homology with a putative phosphate permease of Neurospora crassa suggested that these receptors might serve transport functions. By expression in Xenopus laevis oocytes and in mammalian cells, we have identified Glvr-1 and Ram-1 as sodium-dependent phosphate symporters. Two-electrode voltage-clamp analysis indicates net cation influx, suggesting that phosphate is transported with excess sodium ions. Phosphate uptake was reduced by > 50% in mouse fibroblasts expressing amphotropic envelope glycoprotein, which binds to Ram-1, indicating that Ram-1 is a major phosphate transporter in these cells. RNA analysis shows wide but distinct tissue distributions, with Glvr-1 expression being highest in bone marrow and Ram-1 in heart. Overexpression of Ram-1 severely repressed Glvr-1 synthesis in fibroblasts, suggesting that transporter expression may be controlled by net phosphate accumulation. Accordingly, depletion of extracellular phosphate increased Ram-1 and Glvr-1 expression 3- to 5-fold. These results suggest simple methods to modulate retroviral receptor expression, with possible applications to human gene therapy.
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Improved retroviral vectors for gene transfer and expression.
A. Dusty Miller, G J Rosman · PubMed · 1989 · 2K citations
A Method for Isolation of Intact, Translationally Active Ribonucleic Acid
Guy Cathala, Jean François Savouret, Bernardita Méndez et al. · DNA · 1983 · 1.5K citations