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. 1982 Jan;2(1):30–41. doi: 10.1128/mcb.2.1.30

Assignment of two mitochondrially synthesized polypeptides to human mitochondrial DNA and their use in the study of intracellular mitochondrial interaction.

N A Oliver, D C Wallace
PMCID: PMC369750  PMID: 6955589

Abstract

Two mitochondrially synthesized marker polypeptides, MV-1 and MV-2, were found in human HeLa and HT1080 cells. These were assigned to the mitochondrial DNA in HeLa-HT1080 cybrids and hybrids by demonstrating their linkage to cytoplasmic genetic markers. These markers include mitochondrial DNA restriction site polymorphisms and resistance to chloramphenicol, an inhibitor of mitochondrial protein synthesis. In the absence of chloramphenicol, the expression of MV-1 and MV-2 in cybrids and hybrids was found to be directly proportional to the ratio of the parental mitochondrial DNAs. In the presence of chloramphenicol, the marker polypeptide linked to the chloramphenicol-sensitive mitochondrial DNA continued to be expressed. This demonstrated that resistant and sensitive mitochondrial DNAs can cooperate within a cell for gene expression and that the CAP-resistant allele was dominant or codominant to sensitive. Such cooperation suggests that mitochondrial DNAs can be exchanged between mitochondria.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Apirion D. Three genes that affect Escherichia coli ribosomes. J Mol Biol. 1967 Dec 14;30(2):255–275. [PubMed] [Google Scholar]
  2. Barrell B. G., Anderson S., Bankier A. T., de Bruijn M. H., Chen E., Coulson A. R., Drouin J., Eperon I. C., Nierlich D. P., Roe B. A. Different pattern of codon recognition by mammalian mitochondrial tRNAs. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3164–3166. doi: 10.1073/pnas.77.6.3164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barrell B. G., Bankier A. T., Drouin J. A different genetic code in human mitochondria. Nature. 1979 Nov 8;282(5735):189–194. doi: 10.1038/282189a0. [DOI] [PubMed] [Google Scholar]
  4. Battey J., Clayton D. A. The transcription map of human mitochondrial DNA implicates transfer RNA excision as a major processing event. J Biol Chem. 1980 Dec 10;255(23):11599–11606. [PubMed] [Google Scholar]
  5. Birky C. W., Jr On the origin of mitochondrial mutants: evidence for intracellular selection of mitochondria in the origin of antibiotic-resistant cells in yeast. Genetics. 1973 Jul;74(3):421–432. doi: 10.1093/genetics/74.3.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bogenhagen D., Clayton D. A. The number of mitochondrial deoxyribonucleic acid genomes in mouse L and human HeLa cells. Quantitative isolation of mitochondrial deoxyribonucleic acid. J Biol Chem. 1974 Dec 25;249(24):7991–7995. [PubMed] [Google Scholar]
  7. Borst P., Grivell L. A. The mitochondrial genome of yeast. Cell. 1978 Nov;15(3):705–723. doi: 10.1016/0092-8674(78)90257-x. [DOI] [PubMed] [Google Scholar]
  8. Bunn C. L., Mitchell C. H., Lukins H. B., Linnane A. W. Biogenesis of mitochondria. 18. A new class of cytoplasmically determined antibiotic resistant mutants in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1233–1240. doi: 10.1073/pnas.67.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Chen T. R. In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Exp Cell Res. 1977 Feb;104(2):255–262. doi: 10.1016/0014-4827(77)90089-1. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Costantino P., Attardi G. Identification of discrete electrophoretic components among the products of mitochondrial protein synthesis in HeLa cells. J Mol Biol. 1975 Aug 5;96(2):291–306. doi: 10.1016/0022-2836(75)90349-6. [DOI] [PubMed] [Google Scholar]
  13. Croce C. M. Loss of mouse chromosomes in somatic cell hybrids between HT-1080 human fibrosarcoma cells and mouse peritioneal macrophages. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3248–3252. doi: 10.1073/pnas.73.9.3248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Doersen C. J., Stanbridge E. J. Cytoplasmic inheritance of erythromycin resistance in human cells. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4549–4553. doi: 10.1073/pnas.76.9.4549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dujon B., Slonimski P. P., Weill L. Mitochondrial genetics IX: A model for recombination and segregation of mitochondrial genomes in saccharomyces cerevisiae. Genetics. 1974 Sep;78(1):415–437. doi: 10.1093/genetics/78.1.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Eperon I. C., Anderson S., Nierlich D. P. Distinctive sequence of human mitochondrial ribosomal RNA genes. Nature. 1980 Jul 31;286(5772):460–467. doi: 10.1038/286460a0. [DOI] [PubMed] [Google Scholar]
  17. Francke U., Oliver N. Quantitative analysis of high-resolution trypsin-giemsa bands on human prometaphase chromosomes. Hum Genet. 1978 Dec 18;45(2):137–165. doi: 10.1007/BF00286957. [DOI] [PubMed] [Google Scholar]
  18. Freeman K. B. Action of the N-trifluoroacetyl analogue of D-chloramphenicol. Antimicrob Agents Chemother. 1977 Mar;11(3):563–565. doi: 10.1128/aac.11.3.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gupta R. S., Siminovitch L. An in vitro analysis of the dominance of emetine sensitivity in Chinese hamster ovary cell hybrids. J Biol Chem. 1978 Jun 10;253(11):3978–3982. [PubMed] [Google Scholar]
  20. Gupta R. S., Siminovitch L. Mutants of CHO cells resistant to the protein synthesis inhibitor emetine: genetic and biochemical characterization of second-step mutants. Somatic Cell Genet. 1978 Jan;4(1):77–94. doi: 10.1007/BF01546494. [DOI] [PubMed] [Google Scholar]
  21. Hare J. F., Ching E., Attardi G. Isolation, subunit composition, and site of synthesis of human cytochrome c oxidase. Biochemistry. 1980 May 13;19(10):2023–2030. doi: 10.1021/bi00551a003. [DOI] [PubMed] [Google Scholar]
  22. Hoffmann H. P., Avers C. J. Mitochondrion of yeast: ultrastructural evidence for one giant, branched organelle per cell. Science. 1973 Aug 24;181(4101):749–751. doi: 10.1126/science.181.4101.749. [DOI] [PubMed] [Google Scholar]
  23. Johnson L. V., Walsh M. L., Chen L. B. Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A. 1980 Feb;77(2):990–994. doi: 10.1073/pnas.77.2.990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kit S., Dubbs D. R., Frearson P. M. HeLa cells resistant to bromodeoxyuridine and deficient in thymidine kinase activity. Int J Cancer. 1966 Jan;1(1):19–30. doi: 10.1002/ijc.2910010105. [DOI] [PubMed] [Google Scholar]
  25. LEDERBERG E. M., CAVALLI-SFORZA L., LEDERBERG J. INTERACTION OF STREPTOMYCIN AND A SUPPRESSOR FOR GALACTOSE FERMENTATION IN E. COLI K-12. Proc Natl Acad Sci U S A. 1964 Apr;51:678–682. doi: 10.1073/pnas.51.4.678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LEDERBERG J. Streptomycin resistance; a genetically recessive mutation. J Bacteriol. 1951 May;61(5):549–550. doi: 10.1128/jb.61.5.549-550.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  30. Lansman R. A., Clayton D. A. Mitochondrial protein synthesis in mouse L-cells: effect of selective nicking of mitochondrial DNA. J Mol Biol. 1975 Dec 25;99(4):777–793. doi: 10.1016/s0022-2836(75)80184-7. [DOI] [PubMed] [Google Scholar]
  31. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  32. Linnane A. W., Saunders G. W., Gingold E. B., Lukins H. B. The biogenesis of mitochondria. V. Cytoplasmic inheritance of erythromycin resistance in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1968 Mar;59(3):903–910. doi: 10.1073/pnas.59.3.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Morais R., Giguère L. On the adaptation of cultured chick embryo cells to growth in the presence of chloramphenicol. J Cell Physiol. 1979 Oct;101(1):77–88. doi: 10.1002/jcp.1041010110. [DOI] [PubMed] [Google Scholar]
  34. Nass M. M. Mitochondrial DNA. I. Intramitochondrial distribution and structural relations of single- and double-length circular DNA. J Mol Biol. 1969 Jun 28;42(3):521–528. doi: 10.1016/0022-2836(69)90240-x. [DOI] [PubMed] [Google Scholar]
  35. Nomura M., Engbaek F. Expression of ribosomal protein genes as analyzed by bacteriophage Mu-induced mutations. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1526–1530. doi: 10.1073/pnas.69.6.1526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ojala D., Merkel C., Gelfand R., Attardi G. The tRNA genes punctuate the reading of genetic information in human mitochondrial DNA. Cell. 1980 Nov;22(2 Pt 2):393–403. doi: 10.1016/0092-8674(80)90350-5. [DOI] [PubMed] [Google Scholar]
  37. Posakony J. W., England J. M., Attardi G. Morphological heterogeneity of HeLa cell mitochondria visualized by a modified diaminobenzidine staining technique. J Cell Sci. 1975 Nov;19(2):315–329. doi: 10.1242/jcs.19.2.315. [DOI] [PubMed] [Google Scholar]
  38. Rasheed S., Nelson-Rees W. A., Toth E. M., Arnstein P., Gardner M. B. Characterization of a newly derived human sarcoma cell line (HT-1080). Cancer. 1974 Apr;33(4):1027–1033. doi: 10.1002/1097-0142(197404)33:4<1027::aid-cncr2820330419>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  39. Smith C. A., Jordan J. M., Vinograd J. In vivo effects of intercalating drugs on the superhelix density of mitochondrial DNA isolated from human and mouse cells in culture. J Mol Biol. 1971 Jul 28;59(2):255–272. doi: 10.1016/0022-2836(71)90050-7. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Wallace D. C. Assignment of the chloramphenicol resistance gene to mitochondrial deoxyribonucleic acid and analysis of its expression in cultured human cells. Mol Cell Biol. 1981 Aug;1(8):697–710. doi: 10.1128/mcb.1.8.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wallace D. C., Bunn C. L., Eisenstadt J. M. Cytoplasmic transfer of chloramphenicol resistance in human tissue culture cells. J Cell Biol. 1975 Oct;67(1):174–188. doi: 10.1083/jcb.67.1.174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wiseman A., Attardi G. Cytoplasmically inherited mutations of a human cell line resulting in deficient mitochondrial protein synthesis. Somatic Cell Genet. 1979 Mar;5(2):241–262. doi: 10.1007/BF01539164. [DOI] [PubMed] [Google Scholar]
  44. Yatscoff R. W., Goldstein S., Freeman K. B. Conservation of genes coding for proteins synthesized in human mitochondria. Somatic Cell Genet. 1978 Nov;4(6):633–645. doi: 10.1007/BF01543155. [DOI] [PubMed] [Google Scholar]
  45. de Jong L., Holtrop M., Kroon A. M. The biogenesis of rat mitochondrial ATPase. Two subunits are synthesized inside the mitochondria. Biochim Biophys Acta. 1980 Jun 27;608(1):32–38. doi: 10.1016/0005-2787(80)90130-6. [DOI] [PubMed] [Google Scholar]
  46. de Jong L., Holtrop M., Kroon A. M. The biogenesis of rat-liver mitochondrial ATPase. Evidence that the N, N'-dicyclohexyl carbodiimide-binding protein is synthesized outside the mitochondria. Biochim Biophys Acta. 1980 Feb 29;606(2):331–337. doi: 10.1016/0005-2787(80)90042-8. [DOI] [PubMed] [Google Scholar]

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