Abstract
This study describes the selection and preliminary characterization of mammalian cells resistant to 100 mug Tevenel/ml. Tevenel, the sulfamoyl analog of chloramphenicol, is a specific inhibitor of mitochondrial protein synthesis. After growth in suspension culture for 5 days in 100 mug Tevenel/ml and subsequent plating in 100 mug Tevenel/ml, LMTK- cells yielded resistant clones. As a control, L cells treated identically yielded no clones. Three resistant clones were chosen for study. Each resistant cell line had an identical growth rate in the presence and absence of 100 mug Tevenel/ml. By plating efficiency analysis, the resistant cells were found to be cross-resistant to D- chloramphenicol. The change responsible for resistance was found to be stable for at least 100 generations in the absence of the drug. Protein synthesis by isolated mitochondria of resistant cells was found to be less inhibited by concentrations of both Tevenel and D-chloramphenicol up to 200 mug/ml than the protein synthesis by LMTK- mitochondria. This resistance in vitro was not changed by incubation of the mitochondria in 0.01% Triton X-100.
Full Text
The Full Text of this article is available as a PDF (442.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Attardi B., Attardi G. Persistence of thymidine kinase activity in mitochondria of a thymidine kinase-deficient derivative of mouse L cells. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2874–2878. doi: 10.1073/pnas.69.10.2874. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Basilico C., Joklik W. K. Studies on a temperature-sensitive mutant of vaccinia virus strain WR. Virology. 1968 Dec;36(4):668–677. doi: 10.1016/0042-6822(68)90198-0. [DOI] [PubMed] [Google Scholar]
- Bunn C. L., Wallace D. C., Eisenstadt J. M. Cytoplasmic inheritance of chloramphenicol resistance in mouse tissue culture cells. Proc Natl Acad Sci U S A. 1974 May;71(5):1681–1685. doi: 10.1073/pnas.71.5.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coen D., Deutsch J., Netter P., Petrochilo E., Slonimski P. P. Mitochondrial genetics. I. Methodology and phenomenology. Symp Soc Exp Biol. 1970;24:449–496. [PubMed] [Google Scholar]
- Coon H. G., Horak I., Dawid I. B. Propagation of both parental mitochondrial DNAs in rat-human and mouse-human hybrid cells. J Mol Biol. 1973 Dec 15;81(3):285–298. doi: 10.1016/0022-2836(73)90142-3. [DOI] [PubMed] [Google Scholar]
- Costantino P., Attardi G. Atypical pattern of utilization of amino acids for mitochondrial protein synthesis in HeLa cells. Proc Natl Acad Sci U S A. 1973 May;70(5):1490–1494. doi: 10.1073/pnas.70.5.1490. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FREEMAN K. B. PROTEIN SYNTHESIS IN MITOCHONDRIA. 4. PREPARATION AND PROPERTIES OF MITOCHONDRIA FROM KREBS II MOUSE ASCITES-TUMOUR CELLS. Biochem J. 1965 Feb;94:494–501. doi: 10.1042/bj0940494. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fettes I. M., Haldar D., Freeman K. B. Effect of chloramphenicol on enzyme synthesis and growth of mammalian cells. Can J Biochem. 1972 Feb;50(2):200–209. doi: 10.1139/o72-027. [DOI] [PubMed] [Google Scholar]
- Freeman K. B. Effects of chloramphenicol and its isomers and analogues on the mitochondrial respiratory chain. Can J Biochem. 1970 Apr;48(4):469–478. doi: 10.1139/o70-076. [DOI] [PubMed] [Google Scholar]
- Freeman K. B., Haldar D. The inhibition of NADH oxidation in mammalian mitochondria by chloramphenicol. Biochem Biophys Res Commun. 1967 Jul 10;28(1):8–12. doi: 10.1016/0006-291x(67)90397-x. [DOI] [PubMed] [Google Scholar]
- Freeman K. B., Haldar D. The inhibition of mammalian mitochondrial NADH oxidation by chloramphenicol and its isomers and analogues. Can J Biochem. 1968 Sep;46(9):1003–1008. doi: 10.1139/o68-151. [DOI] [PubMed] [Google Scholar]
- Freeman K. B. Inhibition of mitochondrial and bacterial protein synthesis by chloramphenicol. Can J Biochem. 1970 Apr;48(4):479–485. doi: 10.1139/o70-077. [DOI] [PubMed] [Google Scholar]
- Haldar D., Freeman K. B. Importance of the osmolarity of the incubation medium on amino acid incorporation into protein by isolated rat liver mitochondria. Biochem J. 1969 Mar;111(5):653–661. doi: 10.1042/bj1110653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haldar D., Freeman K. B. The inhibition of protein synthesis and respiration in mouse ascites tumor cells by chloramphenicol and its isomers and analogues. Can J Biochem. 1968 Sep;46(9):1009–1017. doi: 10.1139/o68-152. [DOI] [PubMed] [Google Scholar]
- KIT S., DUBBS D. R., PIEKARSKI L. J., HSU T. C. DELETION OF THYMIDINE KINASE ACTIVITY FROM L CELLS RESISTANT TO BROMODEOXYURIDINE. Exp Cell Res. 1963 Aug;31:297–312. doi: 10.1016/0014-4827(63)90007-7. [DOI] [PubMed] [Google Scholar]
- Kislev N., Spolsky C. M., Eisenstadt J. M. Effect of chloramphenicol on the ultrastructure of mitochondria in sensitive and resistant strains of HeLa. J Cell Biol. 1973 May;57(2):571–579. doi: 10.1083/jcb.57.2.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kit S., Leung W. C., Trkula D. Distinctive properties of mitochondrial thymidine(dT)kinase from bromodeoxyuridine(dBU)-resistant mouse lines. Biochem Biophys Res Commun. 1973 Sep 5;54(1):455–461. doi: 10.1016/0006-291x(73)90943-1. [DOI] [PubMed] [Google Scholar]
- Kit S., Leung W. C., Trkula D. Properties of mitochondrial thymidine kinases of parental and enzyme-deficient HeLa cells. Arch Biochem Biophys. 1973 Oct;158(2):503–513. doi: 10.1016/0003-9861(73)90542-0. [DOI] [PubMed] [Google Scholar]
- Klietmann W., Sato N., Nass M. M. Establishment and characterization of ethidium bromide resistance in simian virus 40-transformed hamster cells. Effects on mitochondria in vivo. J Cell Biol. 1973 Jul;58(1):11–26. doi: 10.1083/jcb.58.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LITMAN R. M., PARDEE A. B. Production of bacteriophage mutants by a disturbance of deoxyribonucleic acid metabolism. Nature. 1956 Sep 8;178(4532):529–531. doi: 10.1038/178529b0. [DOI] [PubMed] [Google Scholar]
- LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Mahler H. R. Biogenetic autonomy of mitochondria. CRC Crit Rev Biochem. 1973 Aug;1(3):381–460. doi: 10.3109/10409237309105439. [DOI] [PubMed] [Google Scholar]
- Molloy P. L., Howell N., Plummer D. T., Linnane A. W., Lukins H. B. Mitochondrial mutants of the yeast Saccharomyces cerevisiae showing resistance in vitro to chloramphenicol inhibition of mitochondrial protein synthesis. Biochem Biophys Res Commun. 1973 May 1;52(1):9–14. doi: 10.1016/0006-291x(73)90946-7. [DOI] [PubMed] [Google Scholar]
- PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rifkin M. R., Luck D. J. Defective production of mitochondrial ribosomes in the poky mutant of Neurospora crassa. Proc Natl Acad Sci U S A. 1971 Feb;68(2):287–290. doi: 10.1073/pnas.68.2.287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sambrook J. F., Padgett B. L., Tomkins J. K. Conditional lethal mutants of rabbitpox virus. I. Isolation of host cell-dependent and temperature-dependent mutants. Virology. 1966 Apr;28(4):592–599. doi: 10.1016/0042-6822(66)90244-3. [DOI] [PubMed] [Google Scholar]
- Spolsky C. M., Eisenstadt J. M. Chloramphenicol-resistant mutants of human HeLa cells. FEBS Lett. 1972 Sep 15;25(2):319–324. doi: 10.1016/0014-5793(72)80514-3. [DOI] [PubMed] [Google Scholar]
- Wallace R. B., Freeman K. B. Initiation of mammalian mitochondrial protein synthesis. The effect of methotrexate. Biochim Biophys Acta. 1974 Nov 6;366(4):466–473. doi: 10.1016/0005-2787(74)90044-6. [DOI] [PubMed] [Google Scholar]
- Yunis A. A., Manyan D. R., Arimura G. K. Comparative effect of chloramphenicol and thiamphenicol on DNA and mitochondrial protein synthesis in mammalian cells. J Lab Clin Med. 1973 May;81(5):713–718. [PubMed] [Google Scholar]