Abstract
The extent to which the cellular processing of shuttle vector-carried genes reflects that of endogenous chromosomal loci has been a subject of considerable controversy. In order to address this issue, we have developed a retroviral-based shuttle vector carrying the Chinese hamster ovary (CHO) adenine phosphoribosyltransferase (aprt) gene stably integrated into the genome to be used for studying mutational specificity in mammalian cells. Initially, we have characterized a collection of UV-induced mutants in a CHO cell background. We have therefore been able to directly compare this shuttle vector data to that previously obtained for UV-induced mutation at the endogenous CHO (aprt)locus. Although some potential differences between the two spectra have been noted, there appears to be a remarkable similarity in the distribution and site specificity of UV-induced mutations. These similarities extend to extrachromosomal shuttle vectors as well and consolidate the role of shuttle vectors as powerful analytical tools for studying mechanisms of point mutagenesis in mammalian cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature cited
Glickman, B.W., de Jong, P.J., de Boer, J.G., Drobetsky, E.A., and Grosovsky, A.J. (1989).Mutat. Res.,226:245–252.
Myers, R.M., Larin, Z., and Maniatis, T. (1985).Science 230:1242–1246.
Fischer, S.G., and Lerraan, L.S. (1983).Proc. Natl. Acad. Sci. U.S.A. 80:1579–1583.
Urlaub, G., Carothers, A.M., and Chasin, L.A. (1985).Proc. Natl. Acad. Sci. U.S.A. 82:1189–1193.
Hauser, J., Seidman, M.M., Sidur, K., and Dixon, K. (1986).Mol. Cell Biol. 6:277–285.
Hsia Chao, H., Lebkowski, J.S., Leong, P., Calos, M.P., and Miller, J.H. (1989).J. Mol. Biol. 205:103–113.
Drinkwater, N.R., and Kleindinst, D.K. (1986).Proc. Natl. Acad. Sci. U.S.A. 83:3402–3406.
Ashman, C.R., Jagadeeswaran, P., and Davidson, R.L. (1986).Proc. Natl. Acad. Sci. U.S.A. 83:3356–3360.
Glazer, P.M., Sarkar, S.N., and Summers, W.C. (1986).Proc. Natl. Acad. Sci. U.S.A. 83:1041–1044.
Drobetsky, E.A., Grosovsky, A.J., and Glickman, B.W. (1987).Proc. Natl. Acad. Sci. U.S.A. 84:9103–9107.
Grosovsky, A.J., deBoer, J.G., deJong, P.J., Drobetsky, E.A., and Glickman, B.W. (1988).Proc. Natl. Acad. Sci. U.S.A. 85:185–188.
Mazur, M.M., and Glickman, B.W. (1988).Somat. Cell Mol. Genet. 14:393–400.
De Boer, J., and Glickman, B.W. (1988).Carcinogenesis (in press).
De Jong, P., Grosovsky, A.J., and Glickman, B.W. (1988).Proc. Natl. Acad. Sci. U.S.A. 85:3499–3505.
Grosovsky, A.J., Drobetsky, E.A., deJong, P.J., and Glickman, B.W. (1986).Genetics 113:405–410.
Cepko, C.L., Roberts, B.E., and Mulligan, R.C. (1984).Cell 37:1053–1062.
Mann, R., Mulligan, R.C., and Baltimore, D. (1983).Cell 37:153–159.
Gluzman, Y. (1981).Cell 23:175–182.
Hirt, B. (1967).J. Mol. Biol. 26:365–369.
Levinson, A., Silver, D., and Seed, B. (1984).J. Mol. Appl. Genet. 2:507–517.
Sanger, F., Nicklen, S., and Coulson, A.R. (1977).Proc. Natl. Acad. Sci. U.S.A. 74:5463–5467.
Scharf, S.J., Horn, G.T., and Erlich, H.A. (1986).Science 233:1076–1078.
Protic-Sabljic, M., N., Tuteja, N. Tuteja, Munson, P.J., Hauser, J., Kraemer, K.H., Dixon, K. (1986).Mol. Cell Biol. 6:3349–3356.
Seidman, M.M., Bredberg, A., Seetharam, S., and Kraemer, K.H. (1987).Proc. Natl. Acad. Sci. U.S.A. 84:4944–4948.
Brash, D.E., Seetharam, S., Kraemer, K.H., Seidman, M.M., and Bredberg, A. (1987).Proc. Natl. Acad. Sci. U.S.A. 84:3782–3786.
Glickman, B.W., Schaaper, R.M., Haseltine, W.A., Dunn, R.L., and Brash, D.E. (1986).Proc. Natl. Acad. Sci. U.S.A. 83:6945–6950.
Schaaper, R.M., Dunn, R.L., and Glickman, B.W. (1987).J. Mol. Biol. 198:187–202.
Franklin, W.A., Doetsch, P.W., and Haseltine, W.A. (1985).Nucleic Acids Res. 13:5317–5325.
Brash, D.E., and Haseltine, W.A. (1982).Nature 298:189–192.
Bohr, V.A., Smith, C.A., Okumoto, D.S., and Hanawalt, P.C. (1985).Cell 40:359–369.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Drobetsky, E.A., Grosovsky, A.J., Skandalis, A. et al. Perspectives on UV light mutagenesis: Investigation of the CHOaprt gene carried on a retroviral shuttle vector. Somat Cell Mol Genet 15, 401–409 (1989). https://doi.org/10.1007/BF01534891
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01534891