Skip to main content
Log in

Selection by [3H] amino acids of CHO-cell mutants with altered leucyl- and asparagyl-transfer RNA synthetases

  • Published:
Somatic Cell Genetics

Abstract

Efficient selection procedures, using [3 H]amino acids as the selecting agent, were developed for isolating temperature-sensitive (TS) mutations in CHO cells affecting protein synthesis. After chemical mutagenesis, leucyl-tRNA synthetase mutants were obtained when [3 H]leucine was used as the selecting agent in two independent experiments. These mutations seem to involve the same genetic locus as the TSH1 mutant described previously (1). A selection with [3 H]valine, in which all amino acids except leucine were at low concentration in the selective medium, resulted in a new class of mutants with reduced asparagyl-tRNA synthetase activity. These results were consistent with the finding that all mutants were phenotypically dependent on the concentration of amino acid, specific to the altered synthetase, in the medium. Our observations suggest that although leucyl synthetase mutations are a relatively common class of TS mutations in CHO cells, the spectrum of mutants obtained can be at least partially manipulated through concentrations of amino acids in selective media. The asparagylsynthetase mutation was shown to be recessive and to complement the leucylsynthetase mutation in cell-cell hybrids.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature cited

  1. Thompson, L. H., Harkins, J. L., and Stanners, C. P. (1973).Proc. Nat. Acad. Sci. USA 70:3094–3098.

    PubMed  Google Scholar 

  2. Thompson, L. H., and Baker, R. M. (1973). InMethods in Cell Biology, (ed.) D. M. Prescott, Vol. 6: pp. 209–281, Academic Press, New York, New York.

    Google Scholar 

  3. Stanners, C. P., and Thompson, L. H. (1974). InControl of Proliferation in Animal Cells, Cold Spring Harbor Conferences on Cell Proliferation, (eds.) Clarkson, B., Baserga, R., Vol. 1: pp. 191–203, Cold Spring Harbor Laboratory.

  4. Puck, T. T., Cieciura, S. J., and Robinson, A. (1958).J. Exp. Med. 108:945–955.

    PubMed  Google Scholar 

  5. McBurney, M., and Whitmore, G. F. (1975).J. Cell Physiol., in press.

  6. Stanners, C. P., Elicieri, G. L., and Green, H. (1971).Nature New Biol. 230:52–54.

    PubMed  Google Scholar 

  7. Baker, R. M., Brunette, D. M., Mankovitz, R., Thompson, L. H., Whitmore, G. F., Siminovitch, L., and Till, J. E. (1974).Cell 1:9–21.

    Google Scholar 

  8. Hampel, A., and Enger, M. D. (1973).J. Mol. Biol. 79:285–293.

    PubMed  Google Scholar 

  9. Molnar, S. J., and Rauth, A. M. (1975).J. Cell. Physiol., in press.

  10. Meiss, H. K., and Basilico, C. (1972).Nature (London) New Biol. 239:66–68.

    Google Scholar 

  11. Scheffler, I. E., and Buttin, G. (1973).J. Cell. Physiol. 81:199–216.

    PubMed  Google Scholar 

  12. Wittes, R. E., and Ozer, H. L. (1973).Exp. Cell Res. 80:127–136.

    PubMed  Google Scholar 

  13. Smith, B. J., and Wigglesworth, N. M. (1972).J. Cell. Physiol. 80:253–258.

    PubMed  Google Scholar 

  14. Roscoe, D. H., Read, M., and Robinson, H. (1973).J. Cell. Physiol. 82:325–332.

    PubMed  Google Scholar 

  15. Liskay, R. M. (1974).J. Cell. Physiol. 84:49–56.

    PubMed  Google Scholar 

  16. Wang, R. J. (1974).Nature 248:76–78.

    PubMed  Google Scholar 

  17. Vaughan, M. H., and Hansen B. S. (1973).J. Biol. Chem. 248:7087–7096.

    PubMed  Google Scholar 

  18. Straus, D. S., and Ames, B. N. (1973).J. Bacteriol. 115:188–197.

    PubMed  Google Scholar 

  19. McBurney, M. W., and Whitmore, G. F. (1974).J. Cell. Physiol. 83:69–74.

    PubMed  Google Scholar 

  20. Deaven, L. L., and Petersen, D. F. (1973).Chromosoma 41:129–144.

    PubMed  Google Scholar 

  21. Patterson, D., F.-T. Kao, and Puck, T. T. (1974).Proc. Natl. Acad. Sci. USA 71:2057–2061.

    PubMed  Google Scholar 

  22. Glaser, D. A., and Wattenburg, W. H. (1966).Ann. N.Y. Acad. Sci. 139:243–257.

    PubMed  Google Scholar 

  23. Raymond, J. D., Couch, J., Glaser, D. A., and Wehr, C. T. (1973).Abstr., Proceedings of the XIII International Congress of Genetics, p. s226.

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Thompson, L.H., Stanners, C.P. & Siminovitch, L. Selection by [3H] amino acids of CHO-cell mutants with altered leucyl- and asparagyl-transfer RNA synthetases. Somat Cell Mol Genet 1, 187–208 (1975). https://doi.org/10.1007/BF01538547

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01538547

Keywords

Navigation