Skip to main content
SpringerLink
Log in

Isolation of multiple classes of mutants of CHO cells resistant to cyclic AMP

  • Published:
Somatic Cell Genetics

Abstract

From mutagenized Chinese hamster ovary (CHO) cells we have isolated, in a single step, 11 independent mutants resistant to the growthinhibitory effects of 8-Brcyclic AMP, cholera toxin, and methylisobutylxanthine. Two major classes and several subclasses of mutants were obtained. Mutants from all classes have a normal doubling time. None of the mutants respond to cyclic AMP treatment with increased flattening and elongation as do the parental cells. Members of the first class have an altered protein kinase activity which has either an increased Ka for cyclic AMP or an absent response to cyclic AMP. Most of those mutations which result in a protein kinase with increased Ka for cyclic AMP (6/11) are dominant in somatic cell hybrids. Those mutations which result in a protein kinase with little or no response to cyclic AMP (3/11) are recessive. Members of the second major class (2/11) have normal levels of basal and cyclic AMP-dependent protein kinase activity. One is recessive and one is dominant by genetic tests. The basis for the defect in this second class of mutants has not been determined.

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

Access this article

Log in via an institution

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. Robison, G. A., Butcher, R. W., and Sutherland, E. W. (1971).Cyclic AMP (Academic Press, New York).

    Google Scholar 

  2. Pastan, I., and Adhya, S. (1976).Bacteriol. Rev. 40:527–551.

    PubMed  Google Scholar 

  3. Johnson, G. S., Friedman, R. M., and Pastan, I. (1971).Proc. Natl. Acad. Sci. U.S.A. 68:425–429.

    PubMed  Google Scholar 

  4. Johnson, G. S., and Pastan, I. (1972).J. Natl. Cancer Inst. 48:1377–1387.

    PubMed  Google Scholar 

  5. Willingham, M. C., and Pastan, I. (1974).J. Cell Biol. 63:288–294.

    PubMed  Google Scholar 

  6. Otten, J., Johnson, G. S., and Pastan, I. (1971).Biochem. Biophys. Res. Commun. 44:1192–1198.

    PubMed  Google Scholar 

  7. Hsie, A. W., and Puck, T. T. (1971).Proc. Natl. Acad. Sci. U.S.A. 68:358–361.

    PubMed  Google Scholar 

  8. Hsie, A. W., Jones, C., and Puck, T. T. (1971).Proc. Natl. Acad. Sci. U.S.A. 68:1648–1652.

    PubMed  Google Scholar 

  9. Van Veen, J., Nooman, K. D., and Roberts, R. M. (1976).Exp. Cell Res. 103:405–413.

    PubMed  Google Scholar 

  10. Hochman, J., Insel, P. A., Bourne, H. R., Coffino, P.,and Tomkins, G. M. (1974).Proc. Natl. Acad. Sci. U.S.A. 72:5051–5055.

    Google Scholar 

  11. Lemaire, I., and Coffino, P. (1977).J. Cell. Physiol. 92:437–446.

    PubMed  Google Scholar 

  12. Steinberg, R. A., Wetters, T., and Coffino, P. (1978).Cell 15:1351–1361.

    PubMed  Google Scholar 

  13. Gutmann, N. S., Rae, P. A., and Schimmer, B. P. (1978).J. Cell. Physiol. 97:451–460.

    PubMed  Google Scholar 

  14. Masui, H., Reid, L. M., and Glans, C. (1978).Exp. Cell Res. 117:219–230.

    PubMed  Google Scholar 

  15. Simantov, R., and Sachs, L. (1975).J. Biol. Chem. 250:3236–3242.

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  17. Stanley, P., Callibot, V., and Siminovitch, L. (1975).Cell 6:121–128.

    PubMed  Google Scholar 

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

    Google Scholar 

  19. Macpherson, I.A., and Montagnier, L. (1964).Virology 23:291–294.

    PubMed  Google Scholar 

  20. 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 

  21. Corbin, J. D., and Reimann, E. M. (1976). InMethods in Enzymology, Vol. 38, (eds.) Hardman, J. G., and O'Malley, B. W. (Academic Press, New York), pp. 287–290.

    Google Scholar 

  22. Evain, D., Gottesman, M. M., Pastan, I., and Anderson, W. (1979).J. Biol. Chem. 254:6931–6937.

    PubMed  Google Scholar 

  23. Gilman, A. G. (1970).Proc. Natl. Acad. Sci. U.S.A. 67:305–312.

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  25. Pontecorvo, G. (1975).Somat. Cell Genet. 1:397–400.

    PubMed  Google Scholar 

  26. Davidson, R. L., and Gerald, P. S. (1976).Somat. Cell Genet. 2:165–176.

    PubMed  Google Scholar 

  27. Davidson, R. L., O'Malley, K. A., and Wheeler, T. B. (1976).Somat. Cell Genet. 2:271–280.

    PubMed  Google Scholar 

  28. O'Malley, K. A., and Davidson, R. L. (1977).Somat. Cell Genet. 3:441–448.

    PubMed  Google Scholar 

  29. Jha, K. K., and Ozer, H. L. (1976).Somat. Cell Genet. 2:215–223.

    PubMed  Google Scholar 

  30. Costa, M., Gerner, E. W., and Russell, D. H. (1976).Biochem. Biophys. Acta 425:246–255.

    PubMed  Google Scholar 

  31. Beavo, J. A., Bechtel, P. J., and Krebs, E. G. (1974). InMethods in Enzymology, Vol. 38, (eds.) Hardman, J. G., and O'Malley, B. W. (Academic Press, New York), pp. 299–308.

    Google Scholar 

  32. Borman, L. S., Dumont, J. N., and Hsie, A. W. (1974).Exp. Cell Res. 91:422–428.

    Google Scholar 

  33. Insel, P. A., Bourne, H. R., Coffino, P., and Tomkins, G. M. (1975).Science 190:896–898.

    PubMed  Google Scholar 

  34. Friedman, D. L., and Chambers, D. A. (1978).Proc. Natl. Acad. Sci. U.S.A. 75:5286–5290.

    PubMed  Google Scholar 

  35. Li, A. P., O'Neill, J. P., Kawashima, K., and Hsie, A. W. (1977).Arch. Biochem. Biophys. 182:181–187.

    PubMed  Google Scholar 

  36. Chasin, M., and Harris, D. N. (1976). InAdvances in Cyclic Nucleotide Research, Vol. 7, (eds.) Greengard, P., and Robison, G. A. (Raven Press, New York), pp. 225–264.

    Google Scholar 

  37. O'Neill, J. P., Li, A. P., and Hsie, A. W. (1977).Biochim. Biophys. Acta 497:35–45.

    PubMed  Google Scholar 

  38. Storrie, B., Puck, T. T., and Wenger, L. (1978).J. Cell. Physiol. 94:69–76.

    PubMed  Google Scholar 

  39. Bradford, M. M. (1976).Anal. Biochem. 72:248–254.

    PubMed  Google Scholar 

  40. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951).J. Biol. Chem. 193:265–275.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, 20205, Bethesda, Maryland

    Michael M. Gottesman, Alphonse LeCam, Maria Bukowski & Ira Pastan

Authors
  1. Michael M. Gottesman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alphonse LeCam
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maria Bukowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ira Pastan
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gottesman, M.M., LeCam, A., Bukowski, M. et al. Isolation of multiple classes of mutants of CHO cells resistant to cyclic AMP. Somat Cell Mol Genet 6, 45–61 (1980). https://doi.org/10.1007/BF01538695

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01538695

Keywords

Search

Navigation