Summary
The human tumor-derived cell line HeLa S3 and nuclear and mitochondrial gene mutants derived from it are resistant to the aminoglycoside antibiotic, paromomycin (PAR). Other carcinoma-derived cells, SV40-transformed cells, and four human diploid fibroblast cell lines are all sensitive to PAR. Sensitivity is dependent on cell density, and at cell numbers greater than 400/cm2 sensitive cells will proliferate in PAR. The resistance to PAR is inherited in a dominant manner in cell-to-cell fusion hybrids, but is not transferred in cytoplast-to-cell fusions. PAR resistance is therefore encoded by a nuclear gene(s). Resistance to PAR is not caused by changes in the response to mitochondrial or cytoplasmic protein synthesis to PAR in vitro. The uptake of PAR is similar in resistant and sensitive cells, and dimethyl sulfoxide does not render resistant cells more sensitive. Thus, HeLa cell PAR resistance is unlike previously reported ribosomal mutations and may derive from differences in the intracellular metabolism of PAR.
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Buchanan, J. H.; Rattan, S. I. S.; Stevens, A., et al. Intracellular accumulation of a fluorescent derivative of paromomycin in human fibroblasts. J. Cell. Biochem. 20:71–80; 1983.
Bunn, C. L.; Eisenstadt, J. M. Cybrid formation in mouse L cells: the influence of cytoplast to cell ratio. Somatic Cell Genet. 3:335–341; 1977.
Bunn, C. L.; Mitchell, D. Paromomycin resistance in human cells. J. Cell Biol. 95:455a; 1982.
Bunn, C. L.; Tarrant, G. M. Limited lifespan in somatic cell hybrids and cybrids. Exp. Cell Res. 127:385–396; 1980.
Carrasco, L. Selective inhibition of translation in transformed cells. FEBS Lett. 110:341–343; 1980.
Coppin-Raynal, E. Ribosomal suppressors and antisuppressors inPodospora anserina: altered susceptibility to paromomycin and relationship between genetic and phenotypic suppression. Biochem. Genet. 19:729–740; 1981.
Coppin-Raynal, E. Ribosomal control of translational fidelity inPodospora anserina: a suppressor and an antisuppressor affecting the paromomycin-induced misreadingin vitro. Curr. Genet. 5:57–63; 1982.
Coppin-Raynal, E.; LeCoze, D. Mutations relieving hypersensitivity to paromomycin caused by ribosomal supprressors inpodospora anserina. Genet. Res. 40:149–164; 1982.
Davey, P. J.; Haslam, J. M.; Linnane, A. W. Biogenesis of mitochondria 12. The effects of aminoglycoside antibiotics on the mitochondrial and cytoplasmic protein-synthesizing system ofSaccharomyces cerevisiae. Arch. Biochem. Biophys. 136:54–64; 1970.
Ege, T.; Reisbig, R. R.; Rogne, S. Enhancement of DNA-mediated gene transfer by inhibitors of autophagic-lysosomal function. Exp. Cell Res. 155:9–16; 1984.
Gorini, L. Streptomycin and misreading of the genetic code. In: Nomura, M.; Tissières, A.; Lengyel, P., eds. Ribosomes. Cold Spring Harbor Laboratory. Cold Spring Harbor, NY: 1974;791–803.
Jiminez, A.; Davies, J. Expression of a transposable antibiotic resistance element inSaccharomyces. Nature 287:869–871; 1980.
Knapp, L. W.; O’Guin, W. M.; Sawyer, R. H., et al. Clonal variations in keratin intermediate filament expression by human somatic cell hybrids. Exp. Cell Res. 156:359–366; 1985.
Kutzleb, R.; Schweyen, R. J.; Kaudewitz, F. Extrachromosomal inheritance of paromomycin resistance inSaccharomyces cerevisiae. Genetic and biochemical characterization of mutants. Mol. Gen. Genet. 125:91–98; 1973.
Li, M.; Tzagaloff, A.; Underbrink-Lyon, K., et al. Identification of the paromomycin-resistance mutation in the 15S rRNA gene of yeast mitochondria. J. Biol. Chem. 257:5921–5928; 1982.
Linnane, A. W.; Nagley, P. Mitochondrial genetics in perspective: the derivation of a genetic and physical map of the yeast mitochondrial genome. Plasmid 1:324–345; 1978.
Lipton, J. H.; McMurray, W. C. Mitochondrial biogenesis in cultured animal cells. I. Effect of chloramphenicol on morphology and mitochondrial respiratory enzymes. Biochim. Biophys. Acta 477:264–272; 1977.
Littlefield, J. W. Selection of hybrids from matings of fibroblastsin vitro and their presumed recombinants. Science 145:709–710; 1964.
Oliver, N. A.; Wallace, D. C. Assignment of mitochondrially synthesized polypeptides to the human mitochondrial DNA and their use in the study of intracellular mitochondrial interaction. Mol. Cell. Biol. 2:30–41; 1982.
Packer, L.; Nolan, J. S.; Katyare, S., et al. Mitochondrial biogenesis in human fibroblasts. J. Bioenerg. Biomembr. 5:81–105; 1973.
Palmer, E.; Wilhelm, J. M. Mistranslation in a eucaryotic organism. Cell 13:329–334; 1978.
Picard-Bennoun, M. Mutations affecting translational fidelity in the eucaryotePodospora anserina: characterization of two ribosomal restrictive mutations. Mol. Gen. Genet. 183:175–180; 1981.
Randsholt, N.; Ibarrondo, F.; Dequard, M., et al. Analysis of revertants of a ribosomal mutation inPodospora anserina: evidence for new ribosomal mutations which confer hypersensitivity to paromomycin. Biochem. Genet. 20:569–584; 1982.
Rattan, S. I. S. Uptake, accumulation and release of paromomycin by human fibroblasts in culture. IRCS Medical Science 10:807–808; 1982.
Tabak, H. F.; Van der Laan, J. C.; Landegent, J. E., et al. Mitochondrially encoded resistance to paromomycin inSaccharomyces cerevisiae: reinvestigation of a controversy. Plasmid 8:261–275; 1982.
Tischfield, J.; Schafer, I. A.; Dickerman, L. H., et al. Lesch-Nyhan syndrome: repository identification nos. GM-2290, 2291, 2292, 2338, 3115, 3116, and 3117. Cytogenet. Cell Genet. 24:199–200; 1979.
Towers, N. R.; Dixon, H.; Kellerman, G. M., et al. Biogenesis of mitochondria 22. The sensitivity of rat liver mitochondria to antibiotics; a phylogenetic difference between a mammalian system and yeast. Arch. Biochem. Biophys. 151;361–369; 1972.
Tulkens, P.; Trouet, A. The uptake and intracellular accumulation of aminoglycoside antibiotics in lysosomes of cultured rat fibroblasts. Biochem. Pharmacol. 27:415–424; 1978.
Wallace, D. C.; Bunn, C. L.; Eisenstadt, J. M. Cytoplasmic transfer of chloramphenicol resistance in human tissue culture cells. J. Cell Biol. 67:174–188; 1975.
White, F. A.; Bunn, C. L. Segregation of mitochondrial DNA in human somatic cell hybrids. Mol. Gen. Genet. 197:453–460; 1984.
Wilhelm, J. M.; Jessop, J. J.; Pettitt, S. E. Aminoglycoside antibiotics and eukaryotic protein synthesis: stimulation of errors in the translation of natural messengers in extracts of cultured human cells. Biochemistry 17:1149–1153; 1978.
Wilhelm, J. M.; Pettitt, S. E.; Jessop, J. J. Aminoglycoside antibiotics and eukaryotic protein synthesis: structure-function relationships in the stimulation of misreading with a wheat embryo system. Biochemistry 17:1143–1149; 1978.
Yatscoff, R. W.; Goldstein, S.; Freeman, K. B. Conservation of genes coding for proteins synthesized in human mitochondria. Somatic Cell. Genet. 4:633–645; 1978.
Zeigler, M. L.; Davidson, R. L. The effect of hexose on chloramphenicol sensitivity and resistance in Chinese hamster cells. J. Cell. Physiol. 98:627–636; 1979.
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This work was supported by National Institutes of Health grant number AG 02664, University of South Carolina Biomedical Research Support grant number S07 RR7160, and by a grant from the Elsa U. Pardee Foundation, all to C. L. B.
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Bunn, C.L., Baron, J.I. & Mitchell, D. Inherent resistance of HeLa cell derivatives to paromomycin. In Vitro Cell Dev Biol 22, 381–386 (1986). https://doi.org/10.1007/BF02623526
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DOI: https://doi.org/10.1007/BF02623526