Abstract
Mutants of S49 mouse lymphoma cells resistant to cytolysis by analogs of cyclic AMP (cAMP) generally have missense mutations in the gene encoding the regulatory subunit of cAMP-dependent protein kinase. We have compared the mutations in 95 spontaneous isolates with those in 60 mutagen-induced isolates by sequence analysis of amplified cDNAs. Twenty-nine single basepair substitutions in 19 codons produced selectable phenotypes. The spontaneous mutant spectrum was dominated by a CpG transition hotspot in the codon for Arg334. This and other nearby CpG sites were found to be methylated in genomic S49 cell DNA by restriction enzyme analyses. Most of the remaining spontaneous mutants had either G-C→C-G or T-A→G-C transversions, which have been associated with damage caused by oxygen radicals. In contrast, the majority of mutants induced with the alkylating mutagens ethyl methanesulfonate (EMS) and N-methyl-N′-nitro-N-nitrosoguanidine had G-C→A-T mutations at non-CpG sites; in addition, EMS induced several A-T→G-C, A-T→T-A, and G-C→T-A substitutions. A single ICR191-induced mutant analyzed had a unique A-T→G-C lesion. A number of spontaneous and mutagen-induced isolates had closely linked double or triple substitutions, and two isolates had tandem triple substitutions.
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Literature Cited
Cooper, D.N., and Youssoufian, H. (1988).Hum. Genet. 78:151–155.
Mohrenweiser, H.W., and Jones, I.M. (1990).Mutat. Res. 231:87–108.
Lindahl, T. (1990).Mutat. Res. 238:305–311.
Lutz, W.K. (1990).Mutat. Res. 238:287–295.
Xiao, W., and Samson, L. (1993).Proc. Natl. Acad. Sci. U.S.A. 90:2117–2121.
Loeb, L.A., and Cheng, K.C. (1990).Mutat. Res. 238:297–304.
Kunkel, T.A. (1992).J. Biol. Chem. 267:18251–18254.
Harwood, J., Tachibana, A., and Meuth, M. (1991).Mol. Cell. Biol. 11:3163–3170.
Zhang, L.-H., Vrieling, H., van Zeeland, A.A., and Jenssen, D. (1992).J. Mol. Biol. 223:627–635.
Phear, G., Armstrong, W., and Meuth, M. (1989).J. Mol. Biol. 209:577–582.
Fearon, E.R., and Jones, P.A. (1992).FASEB J. 6:2783–2790.
Sommer, S.S. (1992).FASEB J. 6:2767–2774.
Murphy, C.S., and Steinberg, R.A. (1985).Somat. Cell Mol. Genet. 11:605–615.
Steinberg, R.A., Gorman, K.B., Øgreid, D., Døskeland, S.O., and Weber, I.T. (1991).J. Biol. Chem. 266:3547–3553.
Ashman, C.R., and Davidson, R.L. (1987).Proc. Natl. Acad. Sci. U.S.A. 84:3354–3358.
Ikehata, H., Akagi, T., Kimura, H., Akasaka, S., and Kato, T. (1989).Mol. Gen. Genet. 219:349–358.
Steinberg, R.A., and Gorman, K.B. (1992).Mol. Cell. Biol. 12:767–772.
Steinberg, R.A., O'Farrell, P.H., Friedrich, U., and Coffino, P. (1977).Cell 10:381–391.
van Daalen Wetters, T., and Coffino, P. (1983).Mol. Cell. Biol. 3:250–256.
Steinberg, R.A., Murphy, C.S., Russell, J.L., and Gorman, K.B. (1987).Somat. Cell Mol. Genet. 13:645–659.
Steinberg, R.A., van Daalen Wetters, T., and Coffino, P. (1978).Cell 15:1351–1361.
Friedrich, U., and Coffino, P. (1977).Proc. Natl. Acad. Sci. U.S.A. 74:679–683.
Clegg, C.H., Correll, L.A., Cadd, G.G., and McKnight, G.S. (1987).J. Biol. Chem. 262:13111–13119.
Shuntoh, H., and Steinberg, R.A. (1991).J. Cell. Physiol. 146:86–93.
Gorman, K.B., and Steinberg, R.A. (1989).BioTechniques 7:326–331.
Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Manual, (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York).
Feinberg, A.P., and Vogelstein, B. (1984).Anal. Biochem. 137:266.
Rideout, W.M., III, Coetzee, G.A., Olumi, A.F., and Jones, P.A. (1990).Science 249:1288–1290.
Tasheva, E.S., and Roufa, D.J. (1993).Somat. Cell Mol. Genet. 19:275–283.
Øgreid, D., Ekanger, R., Suva, R.H., Miller, J.P., and Døskeland, S.O. (1989).Eur. J. Biochem. 181:19–31.
Perutz, M.F. (1990).J. Mol. Biol. 213:203–206.
Topal, M.D. (1988).Carcinogenesis 9:691–696.
Yang, J.-L., Hu, M.-C., and Wu, C.-W. (1991).J. Mol. Biol. 221:421–430.
Zhang, L.-H., and Jenssen, D. (1991).Carcinogenesis 12:1903–1909.
Preston, B.D., Singer, B., and Loeb, L.A. (1986).Proc. Natl. Acad. Sci. U.S.A. 83:8501–8505.
Eckert, K.A., Ingle, C.A., Klinedinst, D.K., and Drinkwater, N.R. (1988).Mol. Carcinogen. 1:50–56.
Lee, G.S.-F., Blonsky, K.S., On, D.L.V., Savage, E.A., Morgan, A.R., and von Borstel, R.C. (1992).J. Mol. Biol. 223:617–626.
Cabral-Neto, J.B., Gentil, A., Caseira-Cabral, R.E., and Sarasin, A. (1992).J. Biol. Chem. 267:19718–19723.
Gentil, A., Cabral-Neto, J.B., Mariage-Samson, R., Margot, A., Imbach, J.L., Rayner, B., and Sarasin, A. (1992).J. Mol. Biol. 227:981–984.
Kamiya, H., Suzuki, M., Komatsu, Y., Miura, H., Kikuchi, K., Sakaguchi, T., Murata, N. Masutani, C., Hanaoka, F., and Ohtsuka, E. (1992).Nucleic Acids Res. 20:4409–4415.
de Oliveira, R.C., Ribeiro, D.T., Nigro, R.G., Di Mascio, P., and Menck, C.F.M. (1992).Nucleic Acids Res. 20:4319–4323.
McBride, T.J., Schneider, J.E., Floyd, R.A., and Loeb, L.A. (1992).Proc. Natl. Acad. Sci. U.S.A. 89:6866–6870.
Cheng, K.C., Cahill, D.S., Kasai, H., Nishimura, S., and Loeb, L.A. (1992).J. Biol. Chem. 267:166–172.
Moraes, E.C., Keyse, S.M., and Tyrrell, R.M. (1990).Carcinogenesis 11:283–293.
Reed, J., and Hutchinson, F. (1987).Mol. Gen. Genet. 208:446–449.
Kohalmi, S.E., and Kunz, B.A. (1988).J. Mol. Biol. 204:561–568.
Pastink, A., Vreeken, C., Nivard, M.J.M., Searles, L.L., and Vogel, E.W. (1989).Genetics 123:123–129.
Ripley, L.S., and Glickman, B.W. (1983).Cold Spring Harbor Symp. Quant. Biol. 47:851–861.
Ellison, K.S., Dogliotti, E., Connors, T.D., Basu, A.K., and Essigmann, J.M. (1989).Proc. Natl. Acad. Sci. U.S.A. 86:8620–8624.
Mitra, G., Pauly, G.T., Kumar, R., Pei, G.K., Hughes, S.H., Moschel, R.C., and Barbacid, M. (1989).Proc. Natl. Acad. Sci. U.S.A. 86:8650–8654.
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Gorman, K.B., Steinberg, R.A. Spectrum of spontaneous missense mutations causing cyclic AMP-resistance phenotypes in cultured S49 mouse lymphoma cells differs markedly from those of mutations induced by alkylating mutagens. Somat Cell Mol Genet 20, 301–311 (1994). https://doi.org/10.1007/BF02254719
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DOI: https://doi.org/10.1007/BF02254719