Somatic Cell Genetics

, Volume 7, Issue 2, pp 235–253 | Cite as

Isolation and characterization of a UV-sensitive hypermutable aphidicolin-resistant Chinese hamster cell line

  • Chia-Cheng Chang
  • John A. Boezi
  • Stephen T. Warren
  • Carol L. K. Sabourin
  • Philip K. Liu
  • Louis Glatzer
  • James E. Trosko


Aphidicolin is a specific inhibitor of DNA polymerase α and blocks DNA synthesis in vivo. The inhibition of purified α-polymerase has been shown to be competitive with dCTP but not with the other three deoxynucleoside triphosphates (dNTPs). In order to study the various roles that the α-polymerase might play in DNA replication and/or repair, we have attempted to isolate Chinese hamster V79 cells that are resistant to aphidicolin. Four resistant mutants were isolated from BrdU-black light- and UV-mutagenized cells. None of the mutants isolated contains an α-polymerase that is resistant, in crude extract measurements, to aphidicolin. Three mutants isolated, however, were found to be resistant to araC. Two mutants tested were found to be sensitive to cytidine and have elevated levels of dCTP or all 4 dNTPs. These results indicate that they are nucleotide pool mutants instead of α-polymerase mutants. One mutant, aphr-4, is characterized by the following: (1) high level of dCTP; (2) thymidine (or CdR, UdR, auxotrophic; (3) sensitive to thymidine (and AdR, GdR); (4) slow-growing; (5) cytidine sensitive; (6) UV sensitive and hypermutable at the ouabain-resistant locus; and (7) a ninefold increase in frequency of chromatid gaps and breaks when cells are exposed to BrdU- containing medium. Revertants of aphr-4 which are partially aphidicolin-resistant and retain the first three characteristics listed above, but not the others, have been isolated. The appearance of this type of revertant indicates that either aphr- 4 or its “revertant” is a double mutant.


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Literature cited

  1. 1.
    Brundret, K.M., Dalziel, W., Hesp, B., Jarvis, J.A.J., and Neidle, S. (1972).J. Chem. Soc. D. Chem. Commun. 1027–1028.Google Scholar
  2. 2.
    Bucknall, R.A., Moores, H., Simms, R., and Hesp, B. (1973).Antimicrob. Ag. Chemother. 4:294–298.Google Scholar
  3. 3.
    Longiaru, M., Ikeda, J.E., Jarkovsky, Z., Horwitz, S.B., and Horwitz, M.S. (1979).Nucleic Acids Res. 6:3369–3386.PubMedGoogle Scholar
  4. 4.
    Habara, A., Kano, K., Nagano, H., Mano, Y., Ikegami, S., and Yamashita, T. (1980).Biochem. Biophys. Res. Commun. 92:8–12.PubMedGoogle Scholar
  5. 5.
    Pedrali-Noy, G., and Spadari, S. (1979).Biochem. Biophys. Res. Commun. 88:1194–1202.PubMedGoogle Scholar
  6. 6.
    McLennan, A.G. (1980).Biochem. Biophys. Res. Commun. 94:116–122.PubMedGoogle Scholar
  7. 7.
    Ikegami, S., Taguchi, T., Ohashi, M., Oguro, M., Nagano, H., and Mano, Y. (1978).Nature 275:458–460.PubMedGoogle Scholar
  8. 8.
    Wist, E. (1980).Experientia 36:405.PubMedGoogle Scholar
  9. 9.
    Hanaoka, F., Kato, H., Ikegami, S., Ohashi, M., and Yamado, M. (1979).Biochem. Biophys. Res. Commun. 87:575–580.PubMedGoogle Scholar
  10. 10.
    Ohashi, M., Tagushi, T., and Ikegami, S. (1978).Biochem. Biophys. Res. Commun. 82:1084–1090.PubMedGoogle Scholar
  11. 11.
    Oguro, M., Suzuki-Hori, C., Nagano, H., Mano, Y., and Ikegami, S. (1979).Eur. J. Biochem. 97:603–607.PubMedGoogle Scholar
  12. 12.
    Wist, E., and Prydz, H. (1979).Nucleic Acids Res. 6:1583–1590.PubMedGoogle Scholar
  13. 13.
    Oguro, M., Shioda, M., Nagano, H., and Mano, Y. (1980).Biochem. Biophys. Res. Commun. 92:13–19.PubMedGoogle Scholar
  14. 14.
    Ikegami, S., Kawada, K., Kimura, Y., and Suzuki, A. (1979).Agric. Biol. Chem. 43:161–166.Google Scholar
  15. 15.
    Berger, N.A., Kurohara, K.K., Petzold, S.J., and Sikorski, G.W. (1979).Biochem. Biophys. Res. Commun. 89:218–225.PubMedGoogle Scholar
  16. 16.
    Ayusawa, D., Iwata, K., Kozu, T., Ikegami, S., and Seno, T. (1979).Biochem. Biophys. Res. Commun. 91:946–954.PubMedGoogle Scholar
  17. 17.
    Nishimura, M., Yasuda, H., Ikegami, S., Ohashi, M., and Yamada, M. (1979).Biochem. Biophys. Res. Commun. 91:939–945.PubMedGoogle Scholar
  18. 18.
    Ford, D.K., and Yerganian, G. (1958).J. Natl. Cancer Inst. 2:393–425.Google Scholar
  19. 19.
    Carter, G. (1979). InDiagnostic Procedures in Veterinary Microbiology, (Charles C Thomas, Springfield, Illinois), pp. 187–197.Google Scholar
  20. 20.
    Eagle, H. (1959).Science 130:432–437.PubMedGoogle Scholar
  21. 21.
    Chu, E.H.Y., Sun, N.C., and Chang, C.C. (1972).Proc. Natl. Acad. Sci. U.S.A. 69:3459–3463.PubMedGoogle Scholar
  22. 22.
    Perry, P., and Wolff, S. (1974).Nature 251:156–158.PubMedGoogle Scholar
  23. 23.
    Chang, C.C., Trosko, J.E., and Akera, T. (1978).Mutat. Res. 51:85–98.PubMedGoogle Scholar
  24. 24.
    Sabourin, C.L.K., Reno, J.M., and Boezi, J.A. (1978).Arch. Biochem. Biophys. 187:96–101.PubMedGoogle Scholar
  25. 25.
    Lindberg, U., and Skoog, L. (1970).Anal. Biochem. 34:152–160.PubMedGoogle Scholar
  26. 26.
    Skoog, L. (1970).Eur. J. Biochem. 17: 202–208.PubMedGoogle Scholar
  27. 27.
    Giles, K.W., and Myers, A. (1965).Nature 206:93.PubMedGoogle Scholar
  28. 28.
    Meuth, M., Trudel, M., and Siminovitch, L. (1979).Somat. Cell Genet. 5:303–318.Google Scholar
  29. 29.
    Robert de Saint Vincent, B., and Buttin, G. (1979).Somat. Cell Genet. 5:67–82.PubMedGoogle Scholar
  30. 30.
    Rogers, A.M., Hill, R., Lehmann, A.R., Arlett, C.F., and Burns, V.W. (1980).Mutat. Res. 69:139–148.PubMedGoogle Scholar
  31. 31.
    Meuth, M., and Green, H. (1974).Cell 3:367–374.PubMedGoogle Scholar
  32. 32.
    Robert de Saint Vincent, B., Dechamps, M., and Buttin, G. (1980).J. Biol. Chem. 255:162–167.PubMedGoogle Scholar
  33. 33.
    Reichard, P., Canellakis, Z.N., and Canellakis, E.S. (1961).J. Biol. Chem. 236:2514–2519.PubMedGoogle Scholar
  34. 34.
    Meuth, M., Aufreiter, E., and Reichard, P. (1976).Eur. J. Biochem. 71:39–43.PubMedGoogle Scholar
  35. 35.
    Moore, E.C., and Hurlbert, R.B. (1966).J. Biol. Chem. 241:4802–4809.PubMedGoogle Scholar
  36. 36.
    Meuth, M., L'Heureux-Huard, N., and Trudel, M. (1979).Proc. Natl. Acad. Sci. U.S.A. 76:6505–6509.PubMedGoogle Scholar
  37. 37.
    Sutherland, G.R., Baker, E., and Seshadri, R.S. (1980).Am. J. Hum. Genet. 32:542–548.PubMedGoogle Scholar
  38. 38.
    Hsu, T.C., and Somers, C.E. (1961).Proc. Natl. Acad. Sci. U.S.A. 47:396–403.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1981

Authors and Affiliations

  • Chia-Cheng Chang
    • 1
  • John A. Boezi
    • 2
  • Stephen T. Warren
    • 1
  • Carol L. K. Sabourin
    • 2
  • Philip K. Liu
    • 1
  • Louis Glatzer
    • 2
  • James E. Trosko
    • 1
  1. 1.Department of Pediatrics and Human DevelopmentMichigan State UniversityEast Lansing
  2. 2.Department of BiochemistryMichigan State UniversityEast Lansing
  3. 3.Department of BiologyUniversity of ToledoToledo

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