Plasma membrane receptors for ecotropic murine retroviruses require a limiting accessory factor.
A retroviral vector was used to express various amounts of the receptor (ecoR) for ecotropic host range murine retroviruses on naturally barren hamster, mink, and human cells. These cells and murine cells were then incubated for 2 h with dilutions of a helper-free ecotropic retrovirus that encodes human growth hormone, and the number of infected cells was later determined by growth hormone-specific immunofluorescence. For all cells under the conditions of these studies, virus adsorption was the limiting step of infection and the cellular capacities for infection were unsaturated either at cell surfaces or at intracellular sites. Thus, infections occurred at low multiplicities of infection per cell and were directly proportional to virus and cell concentrations, and only a small percentage (ca. 5%) of the infectious virions became adsorbed from the medium during the 2-h incubations. Although increasing the adsorption by raising virus or cell concentrations results in more infections in the cultures, increasing adsorption by raising the number of ecoR above a low threshold had no effect on infections. Thus, cells with a low number of ecoR were infected as efficiently as highly adsorbing cells that contained many times more ecoR. To reconcile these results, we conclude that only a small, set number of cell surface ecoR can be functional for infection and that all excess ecoR can only bind virus into an unsalvageable pool. Therefore, retroviral receptors on single cells are functionally diverse. Our results suggest that activity of ecoR in infection requires a limiting second cellular component.
Wang
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Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health Sciences University, Portland 97201-3098.
Paul
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Burgeson
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Keene
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DR
Kabat
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D016475:3T3 Cells; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006023:Glycoproteins / Q000378:metabolism; D013006:Growth Hormone / Q000235:genetics / Q000502:physiology*; D006801:Humans; D051379:Mice; D016253:Microscopy, Immunoelectron; D011991:Receptors, Virus / Q000235:genetics / Q000502:physiology*; D015183:Restriction Mapping; D012190:Retroviridae / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure
D006023:Glycoproteins; D011991:Receptors, Virus; D013006:Growth Hormone