Abstract
An erythromycin-resistant mutant, ERY2301, was isolated from ethidium bromide-treated HeLa cells in the presence of erythromycin at 300 micrograms/ml. ERY2301 cells were enucleated and the anucleate cytoplasts were fused with D98/AH-2, a hypoxanthine phosphoribosyltransferase-deficient variant of HeLa cells. The resultant cybrids were isolated in a double selective medium containing erythromycin and 6-thioguanine. Cybrid formation occurred at a frequency of 10(-3) to 10(-4). In vitro protein synthesis by intact and Triton X-100 treated mitochondria isolated from ERY2301 was resistant to the macrolide antibiotics erythromycin and carbomycin, but was sensitive to chloramphenicol. These results suggest that the site of erythromycin resistance in ERY2301 may be at the level of mitochondrial protein synthesis and indicate that this trait is cytoplasmically inherited and, therefore, presumably encoded in the mitochondrial genome.
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