Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1979 Dec;76(12):6505–6509. doi: 10.1073/pnas.76.12.6505

Characterization of a mutator gene in Chinese hamster ovary cells.

M Meuth, N L'Heureux-Huard, M Trudel
PMCID: PMC411894  PMID: 293738

Abstract

We have recently reported the isolation of a class of mutants (called thy-) that is both resistant to arabinosyl cytosine and auxotrophic for thymidine. thy- mutants have a 5- to 10-fold elevated pool of dCTP and are deficient in the synthesis of dTTP as an apparent consequence of a single mutation in the gene for ribonucleoside-diphosphate reductase (2'-deoxyribonucleoside-diphosphate:oxidized-thioredoxin 2'-oxidoreductase, EC 1.17.4.1). Here we show that three independent thy- lines have a 5- to 50-fold higher frequency and rate of spontaneous mutation for two genetic markers, 6-thioguanine resistance and ouabain resistance. The higher rate of mutation is site specific because two other genetic markers, reversion of proline auxotrophy to proline prototrophy and emetine resistance, are unaffected. Ouabain- and 6-thioguanine-resistant mutations occur at a much lower rate in revertants of thy- to the wild-type state, so the increased rate of mutation is the consequence of the thy- mutation. Both the increased mutational rate and the increased intracellular pools of dCTP are dominant or codominant in hybrid cells, and alterations of the ratio of the pools of dCTP to dTTP in thy- 49 produce corresponding changes in the rate of mutation. Thus, thy- is a mutator gene in Chinese hamster ovary cells, apparently as a consequence of the imbalance of deoxynucleoside triphosphate pools created by the expanded pool of dCTP.

Full text

PDF
6505

Selected References

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

  1. Bernstein C., Bernstein H., Mufti S., Strom B. Stimulation of mutation in phage T 4 by lesions in gene 32 and by thymidine imbalance. Mutat Res. 1972 Oct;16(2):113–119. doi: 10.1016/0027-5107(72)90171-6. [DOI] [PubMed] [Google Scholar]
  2. Bjursell G., Reichard P. Effects of thymidine on deoxyribonucleoside triphosphate pools and deoxyribonucleic acid synthesis in Chinese hamster ovary cells. J Biol Chem. 1973 Jun 10;248(11):3904–3909. [PubMed] [Google Scholar]
  3. Boersma D., McGill S., Mollenkamp J., Roufa D. J. Emetine resistance in Chinese hamster cells. Analysis of ribosomal proteins prepared from mutant cells. J Biol Chem. 1979 Jan 25;254(2):559–567. [PubMed] [Google Scholar]
  4. Bradley M. O., Sharkey N. A. Mutagenicity of thymidine to cultured Chinese hamster cells. Nature. 1978 Aug 10;274(5671):607–608. doi: 10.1038/274607a0. [DOI] [PubMed] [Google Scholar]
  5. Bridges B. A., Law J., Munson R. J. Mutagenesis in Escherichia coli. II. Evidence for a common pathway for mutagenesis by ultraviolet light, ionizing radiation and thymine deprivation. Mol Gen Genet. 1968;103(3):266–273. doi: 10.1007/BF00273698. [DOI] [PubMed] [Google Scholar]
  6. Caskey C. T., Kruh G. D. The HPRT locus. Cell. 1979 Jan;16(1):1–9. doi: 10.1016/0092-8674(79)90182-x. [DOI] [PubMed] [Google Scholar]
  7. Chasin L. A., Urlaub G. Mutant alleles for hypoxanthine phosphoriboxyltransferase: codominant expression, complementation, and segregation in hybrid Chinese hamster cells. Somatic Cell Genet. 1976 Sep;2(5):453–467. doi: 10.1007/BF01542725. [DOI] [PubMed] [Google Scholar]
  8. Gupta R. S., Siminovitch L. The isolation and preliminary characterization of somatic cell mutants resistant to the protein synthesis inhibitor-emetine. Cell. 1976 Oct;9(2):213–219. doi: 10.1016/0092-8674(76)90112-4. [DOI] [PubMed] [Google Scholar]
  9. Gupta R. S., Siminovitch L. The molecular basis of emetine resistance in Chinese hamster ovary cells: alteration in the 40S ribosomal subunit. Cell. 1977 Jan;10(1):61–66. doi: 10.1016/0092-8674(77)90140-4. [DOI] [PubMed] [Google Scholar]
  10. Kao F. T., Puck T. T. Genetics of somatic mammalian cells. IV. Properties of Chinese hamster cell mutants with respect to the requirement for proline. Genetics. 1967 Mar;55(3):513–524. doi: 10.1093/genetics/55.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaufman E. R., Davidson R. L. Bromodeoxyuridine mutagenesis in mammalian cells: mutagenesis is independent of the amount of bromouracil in DNA. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4982–4986. doi: 10.1073/pnas.75.10.4982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lindberg U., Skoog L. A method for the determination of dATP and dTTP in picomole amounts. Anal Biochem. 1970 Mar;34:152–160. doi: 10.1016/0003-2697(70)90096-5. [DOI] [PubMed] [Google Scholar]
  13. Luria S. E., Delbrück M. Mutations of Bacteria from Virus Sensitivity to Virus Resistance. Genetics. 1943 Nov;28(6):491–511. doi: 10.1093/genetics/28.6.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Meuth M., Aufreiter E., Reichard P. Deoxyribonucleotide pools in mouse-fibroblast cell lines with altered ribonucleotide reductase. Eur J Biochem. 1976 Dec;71(1):39–43. doi: 10.1111/j.1432-1033.1976.tb11087.x. [DOI] [PubMed] [Google Scholar]
  15. Meuth M., Green H. Induction of a deoxycytidineless state in cultured mammalian cells by bromodeoxyuridine. Cell. 1974 Jun;2(2):109–112. doi: 10.1016/0092-8674(74)90099-3. [DOI] [PubMed] [Google Scholar]
  16. Moore E. C., Hurlbert R. B. Regulation of mammalian deoxyribonucleotide biosynthesis by nucleotides as activators and inhibitors. J Biol Chem. 1966 Oct 25;241(20):4802–4809. [PubMed] [Google Scholar]
  17. Nordenskjöld B. A., Skoog L., Brown N. C., Reichard P. Deoxyribonucleotide pools and deoxyribonucleic acid synthesis in cultured mouse embryo cells. J Biol Chem. 1970 Oct 25;245(20):5360–5368. [PubMed] [Google Scholar]
  18. Reichard P. From deoxynucleotides to DNA synthesis. Fed Proc. 1978 Jan;37(1):9–14. [PubMed] [Google Scholar]
  19. SZYBALSKI W., SZYBALSKA E. H. Drug sensitivity as a genetic marker for human cell lines. Med Bull (Ann Arbor) 1962 Sep-Oct;28:277–293. [PubMed] [Google Scholar]
  20. Siminovitch L. On the nature of hereditable variation in cultured somatic cells. Cell. 1976 Jan;7(1):1–11. doi: 10.1016/0092-8674(76)90249-x. [DOI] [PubMed] [Google Scholar]
  21. Skoog L. An enzymatic method for the determination of dCTP and dGTP in picomole amounts. Eur J Biochem. 1970 Dec;17(2):202–208. doi: 10.1111/j.1432-1033.1970.tb01154.x. [DOI] [PubMed] [Google Scholar]
  22. Skoog L., Nordenskjöld B. Effects of hydroxyurea and 1-beta-D-arabinofuranosyl-cytosine on deoxyribonucleotide pools in mouse embryo cells. Eur J Biochem. 1971 Mar 1;19(1):81–89. doi: 10.1111/j.1432-1033.1971.tb01290.x. [DOI] [PubMed] [Google Scholar]
  23. Smith M. D., Green R. R., Ripley L. S., Drake J. W. Thymineless mutagenesis in bacteriophage T4. Genetics. 1973 Jul;74(3):393–403. doi: 10.1093/genetics/74.3.393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. WATSON J. D., CRICK F. H. Genetical implications of the structure of deoxyribonucleic acid. Nature. 1953 May 30;171(4361):964–967. doi: 10.1038/171964b0. [DOI] [PubMed] [Google Scholar]
  25. WATSON J. D., CRICK F. H. The structure of DNA. Cold Spring Harb Symp Quant Biol. 1953;18:123–131. doi: 10.1101/sqb.1953.018.01.020. [DOI] [PubMed] [Google Scholar]
  26. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Worton R. G., Ho C. C., Duff C. Chromosome stability in CHO cells. Somatic Cell Genet. 1977 Jan;3(1):27–45. doi: 10.1007/BF01550985. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES