Comparing the Frequency and Spectra of Mutations Induced When an SV40-Based Shuttle Vector Containing Covalently Bound Residues of Structurally-Related Carcinogens Replicates in Human Cells
It is widely recognized that the transformation of normal cells into malignant cells is a multistep process, and there is substantial evidence that mutations and gene rearrangements are involved in causing one or more of the required changes. However, the molecular mechanisms by which carcinogens induce such changes in mammalian cells are not well understood. One reason for this has been the difficulty involved in sequencing the newly mutated genes to determine the kinds of changes that have taken place. The recent development of shuttle vectors, i.e., small circular plasmid DNA molecules that are capable of replicating in mammalian cells and also in bacteria, has greatly reduced the level of difficulty (Calos et al.,1983; Sarkar et al., 1984; Seidman et al., 1985). It is now feasible to investigate at the DNA sequence level the nature of mutations formed when carcinogen-treated plasmids replicate in human cells.
KeywordsBase Substitution Shuttle Vector Bulky Adduct Chinese Hamster Ovary Cell Mutation Pyrene Diol
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- Beland, F. A., Ribovich, M., Howard, P. C., Heflich, R. H., Kurian, P., and Milo G. E. (1986). Cytotoxicity, cellular transformation and DNA adducts in normal human diploid fibroblasts exposed to 1-nitrosopyrene, a reduced derivative of the environmental contaminant, 1-nitropyrene. Carcinogenesis 7, 1279–1283.CrossRefPubMedGoogle Scholar
- Kraemer, K. H., and Seidman, M. M. (1989). Use of supF, an Escherichia coli tyrosine suppressor tRNA gene, as a mutagenic target in shuttle-vector plasmids. Mutat. Res. 220, 61–72.Google Scholar
- Norman, D., Abuaf, P., Hingerty, B. E., Live, D., Grunberger, D., Broyde, S., and Patel, D. J. (1989). NMR and computational characterization of the N-(deoxyguanosin- 8-yl)aminofluorene adduct RAF)G] opposite adenosine in DNA: (AF)G[syrd-A[anti] pair formation and its pH dependence. Biochemistry 28, 7462–7476.Google Scholar
- Sarkar, S., Dasgupta, U. B., and Summers, W. C. (1984). Error-prone mutagenesis detected in mammalian cells by a shuttle vector containing the supF gene of Escherichia coli. Mol. Cell. Biol. 4, 2227–2230.Google Scholar
- Weinstein, I. B., Jeffrey, A. M., Jennette, K. W., Blobstein, S. H., Harvey, R. G., Harris, C., Autrup, H., Kasai, H., and Nakanishi K. (1976). Benzo[alpyrene diol epoxides as intermediates in nucleic acid binding in vitro and in vivo. Sci. 193, 592–595.Google Scholar
- Yang, J.-L., Maher, V. M., and McCormick, J. J. (1987a). Kinds of mutations formed when a shuttle vector containing adducts of benzo[a]pyrene-7,8-dio1–9,10-epoxide replicates in COS7 cells. Mol. Cell. Biol. 7, 1267–1270.Google Scholar