Abstract
We have investigated the kinds of mutations induced when a shuttle vector containing covalently bound residues of (+/-)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9, 10-tetrahydrobenzo[a]pyrene (BPDE) replicates in human cells. A human embryonic kidney cell line, 293, was used as the eukaryotic host. The target gene for mutation analysis, supF, codes for a tyrosine suppressor tRNA and is strategically located between the origin of replication of the plasmid in Escherichia coli and the gene for a selectable marker, so that the possibility of recovering supF mutants containing gross rearrangements is low. The frequency of supF mutants obtained when untreated plasmid replicated in 293 cells was 1.4 X 10(-4). The frequency with BPDE-treated plasmid increased linearly as a function of the number of adducts, with 16 adducts per plasmid giving 38 X 10(-4). Polyacrylamide gel and agarose gel electrophoresis analysis of 137 plasmids with mutations in the supF gene indicated that 70% (21/30) from untreated plasmids contained deletions or insertions or showed altered gel mobility, whereas only 28% (30/107) of those derived from BPDE-treated plasmids contained such alterations. Of the 86 unequivocally independent mutants derived from BPDE-treated plasmids that were analyzed by sequencing, the majority (60/86) exhibited base substitutions. Mutants exhibiting frameshifts (insertions or deletions of one, two, or four base pairs) were also found, but they were a minority (11/86). In the progeny of BPDE-treated plasmids 61/71 base substitutions observed were transversions, with 45/61 G X C----T X A. Examination of the location of BPDE-induced mutations among the 85 base pairs in the structure of the tRNA revealed that 30% of the base substitutions occurred at two sites and 44% of the rest occurred at five other hot spots. Only 20% of all these base changes involved a site in which a guanine containing a BPDE adduct is predicted to be labile--i.e., a guanine that has a pyrimidine to its 5' side.
Full text
PDF




Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ashman C. R., Davidson R. L. High spontaneous mutation frequency in shuttle vector sequences recovered from mammalian cellular DNA. Mol Cell Biol. 1984 Nov;4(11):2266–2272. doi: 10.1128/mcb.4.11.2266. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aust A. E., Drinkwater N. R., Debien K., Maher V. M., McCormick J. J. Comparison of the frequency of diphtheria toxin and thioguanine resistance induced by a series of carcinogens to analyze their mutational specificities in diploid human fibroblasts. Mutat Res. 1984 Jan;125(1):95–104. doi: 10.1016/0027-5107(84)90036-8. [DOI] [PubMed] [Google Scholar]
- Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bredberg A., Kraemer K. H., Seidman M. M. Restricted ultraviolet mutational spectrum in a shuttle vector propagated in xeroderma pigmentosum cells. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8273–8277. doi: 10.1073/pnas.83.21.8273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Calos M. P., Lebkowski J. S., Botchan M. R. High mutation frequency in DNA transfected into mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3015–3019. doi: 10.1073/pnas.80.10.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Celis J. E., Piper P. W. Compilation of mutant suppressor tRNA sequences. Nucleic Acids Res. 1982 Jan 22;10(2):r83–r91. doi: 10.1093/nar/10.2.762-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chakrabarti S., Joffe S., Seidman M. M. Recombination and deletion of sequences in shuttle vector plasmids in mammalian cells. Mol Cell Biol. 1985 Sep;5(9):2265–2271. doi: 10.1128/mcb.5.9.2265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chakrabarti S., Mizusawa H., Seidman M. Survival and mutagenesis of bacterial plasmids with localized carcinogen adducts. Mutat Res. 1984 Apr;126(2):127–137. doi: 10.1016/0027-5107(84)90054-x. [DOI] [PubMed] [Google Scholar]
- Drinkwater N. R., Klinedinst D. K. Chemically induced mutagenesis in a shuttle vector with a low-background mutant frequency. Proc Natl Acad Sci U S A. 1986 May;83(10):3402–3406. doi: 10.1073/pnas.83.10.3402. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eisenstadt E., Warren A. J., Porter J., Atkins D., Miller J. H. Carcinogenic epoxides of benzo[a]pyrene and cyclopenta[cd]pyrene induce base substitutions via specific transversions. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1945–1949. doi: 10.1073/pnas.79.6.1945. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glazer P. M., Sarkar S. N., Summers W. C. Detection and analysis of UV-induced mutations in mammalian cell DNA using a lambda phage shuttle vector. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1041–1044. doi: 10.1073/pnas.83.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
- Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
- Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
- Hauser J., Seidman M. M., Sidur K., Dixon K. Sequence specificity of point mutations induced during passage of a UV-irradiated shuttle vector plasmid in monkey cells. Mol Cell Biol. 1986 Jan;6(1):277–285. doi: 10.1128/mcb.6.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
- King H. W., Brookes P. On the nature of the mutations induced by the diolepoxide of benzo[a]pyrene in mammalian cells. Carcinogenesis. 1984 Jul;5(7):965–970. doi: 10.1093/carcin/5.7.965. [DOI] [PubMed] [Google Scholar]
- Lebkowski J. S., Clancy S., Miller J. H., Calos M. P. The lacI shuttle: rapid analysis of the mutagenic specificity of ultraviolet light in human cells. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8606–8610. doi: 10.1073/pnas.82.24.8606. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lebkowski J. S., DuBridge R. B., Antell E. A., Greisen K. S., Calos M. P. Transfected DNA is mutated in monkey, mouse, and human cells. Mol Cell Biol. 1984 Oct;4(10):1951–1960. doi: 10.1128/mcb.4.10.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lebkowski J. S., Miller J. H., Calos M. P. Determination of DNA sequence changes induced by ethyl methanesulfonate in human cells, using a shuttle vector system. Mol Cell Biol. 1986 May;6(5):1838–1842. doi: 10.1128/mcb.6.5.1838. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lobanenkov V. V., Plumb M., Goodwin G. H., Grover P. L. The effect of neighbouring bases on G-specific DNA cleavage mediated by treatment with the anti-diol epoxide of benzo[a]pyrene in vitro. Carcinogenesis. 1986 Oct;7(10):1689–1695. doi: 10.1093/carcin/7.10.1689. [DOI] [PubMed] [Google Scholar]
- Loeb L. A., Preston B. D. Mutagenesis by apurinic/apyrimidinic sites. Annu Rev Genet. 1986;20:201–230. doi: 10.1146/annurev.ge.20.120186.001221. [DOI] [PubMed] [Google Scholar]
- Miller J. H., Lebkowski J. S., Greisen K. S., Calos M. P. Specificity of mutations induced in transfected DNA by mammalian cells. EMBO J. 1984 Dec 20;3(13):3117–3121. doi: 10.1002/j.1460-2075.1984.tb02267.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mizusawa H., Lee C. H., Kakefuda T. Alteration of plasmid DNA-mediated transformation and mutation induced by covalent binding of benzo[alpha]pyrene-7,8-dihydrodiol-9,10-oxide in Escherichia coli. Mutat Res. 1981 Jun;82(1):47–57. doi: 10.1016/0027-5107(81)90137-8. [DOI] [PubMed] [Google Scholar]
- Mizusawa H., Lee C. H., Kakefuda T., McKenney K., Shimatake H., Rosenberg M. Base insertion and deletion mutations induced in an Escherichia coli plasmid by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6817–6820. doi: 10.1073/pnas.78.11.6817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osborne M. R., Harvey R. G., Brookes P. The reaction of trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene with DNA involves attack at the N7-position of guanine moieties. Chem Biol Interact. 1978 Jan;20(1):123–130. doi: 10.1016/0009-2797(78)90087-x. [DOI] [PubMed] [Google Scholar]
- Osborne M. R., Jacobs S., Harvey R. G., Brookes P. Minor products from the reaction of (+) and (-) benzo[a]-pyrene-anti-diolepoxide with DNA. Carcinogenesis. 1981;2(6):553–558. doi: 10.1093/carcin/2.6.553. [DOI] [PubMed] [Google Scholar]
- Osborne M., Merrifield K. Depurination of benzo[a]pyrene-diolepoxide treated DNA. Chem Biol Interact. 1985 Feb-Apr;53(1-2):183–195. doi: 10.1016/s0009-2797(85)80095-8. [DOI] [PubMed] [Google Scholar]
- Razzaque A., Chakrabarti S., Joffee S., Seidman M. Mutagenesis of a shuttle vector plasmid in mammalian cells. Mol Cell Biol. 1984 Mar;4(3):435–441. doi: 10.1128/mcb.4.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Razzaque A., Mizusawa H., Seidman M. M. Rearrangement and mutagenesis of a shuttle vector plasmid after passage in mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3010–3014. doi: 10.1073/pnas.80.10.3010. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sarkar S., Dasgupta U. B., Summers W. C. Error-prone mutagenesis detected in mammalian cells by a shuttle vector containing the supF gene of Escherichia coli. Mol Cell Biol. 1984 Oct;4(10):2227–2230. doi: 10.1128/mcb.4.10.2227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seidman M. M., Dixon K., Razzaque A., Zagursky R. J., Berman M. L. A shuttle vector plasmid for studying carcinogen-induced point mutations in mammalian cells. Gene. 1985;38(1-3):233–237. doi: 10.1016/0378-1119(85)90222-7. [DOI] [PubMed] [Google Scholar]
- Strauss B., Rabkin S., Sagher D., Moore P. The role of DNA polymerase in base substitution mutagenesis on non-instructional templates. Biochimie. 1982 Aug-Sep;64(8-9):829–838. doi: 10.1016/s0300-9084(82)80138-7. [DOI] [PubMed] [Google Scholar]
- Topal M. D., Fresco J. R. Complementary base pairing and the origin of substitution mutations. Nature. 1976 Sep 23;263(5575):285–289. doi: 10.1038/263285a0. [DOI] [PubMed] [Google Scholar]
- Weinstein I. B., Jeffrey A. M., Jennette K. W., Blobstein S. H., Harvey R. G., Harris C., Autrup H., Kasai H., Nakanishi K. Benzo(a)pyrene diol epoxides as intermediates in nucleic acid binding in vitro and in vivo. Science. 1976 Aug 13;193(4253):592–595. doi: 10.1126/science.959820. [DOI] [PubMed] [Google Scholar]
- Yang J. L., Maher V. M., McCormick J. J. Kinds of mutations formed when a shuttle vector containing adducts of benzo[a]pyrene-7,8-diol-9,10-epoxide replicates in COS7 cells. Mol Cell Biol. 1987 Mar;7(3):1267–1270. doi: 10.1128/mcb.7.3.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]