Abstract
The hydroxyurea-resistant Chinese hamster cell line 600H has been shown to have greatly elevated quantities of ribonucleotide reductase. This increase in enzyme activity is due to an increased level of both the M1 and M2 subunit activities. The M1 subunit has been purified from the 600H cell line and shown to consist of a series of six protein spots with apparent molecular weights of 88,000 daltons, but with varying isoelectric points in the range of pH 6.5–7.0. Western blot analyses with antisera against the M1 and M2 proteins indicated that both subunit proteins are present in elevated quantities in the 600H cell line when compared to the wild-type V79 cell line. Southern blot analyses with genomic DNA from the series of stepwise-selected hydroxyurea-resistant cell lines leading to 600H showed that, in latter steps of selection, genomic sequences homologous to a mouse M1 cDNA have undergone a fivefold amplication. This was accompanied by a four-to eightfold increase in the single M1 homologous mRNA.
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Literature cited
Thelander, L., and Reichard, P. (1979).Annu. Rev. Biochem. 48:133–158.
Eriksson, S., Gudas, L., Clift, S., Caras, I., Ullman, B., and Martin Jr., D. (1981).J. Biol. Chem. 256:10193–10197.
Mattaliano, R., Sloan, A., Plumer, E., and Klippenstein, G. (1981).Biochem. Biophys. Res. Commun. 102:667–674.
Engström, Y., Eriksson, S., Thelander, L., and Åkerman, M. (1979).Biochemistry 18:2941–2947.
Eriksson, S., Thelander, L., and Akerman, M. (1979).Biochemistry 18:2948–2952.
Caras, I., and Martin Jr., D. (1982).J. Biol. Chem. 257:9508–9512.
Thelander, M., Gräslund, A., and Thelander, L. (1985).J. Biol. Chem. 260:2737–2741.
Meuth, M., and Green, H. (1974).Cell 3:367–374.
Ullman, B., Gudas, L., Caras, I., Eriksson, S., Weinberg, G., Wormsted, M., and Martin, D., Jr. (1981).J. Biol. Chem. 256:10189–10192.
Lewis, W., and Wright, J. (1979).Somat. Cell Genet. 5:83–96.
Åkerblom, L., Ehrenberg, A., Gräslund, A., Lankinen, H., Reichard, P., and Thelander, L. (1981).Proc. Natl. Acad. Sci. U.S.A. 78:2159–2163.
Sabourin, C., Bates, P., Glatzer, L., Chang, C.-C., Trosko, J., and Boezi, J. (1981).Somat. Gell Genet. 7:255–268.
Lewis, W., and Srinivasan, P. (1983).Mol. Cell Biol. 3:1053–1061.
Thelander, L., and Berg, P. (1986).Mol. Cell. Biol. 6:3433–3442.
McClarty, G., Chan, A., and Wright, J. (1986).Somat. Cell. Mol. Genet. 12:121–131.
Wright, J., Alam, T., McClarty, G., Taggert, A., and Thelander, L. (1987).Somat. Cell Mol. Genet. (in press).
Thirion, J.-P., Banville, D., and Noel, H. (1976).Genetics 83:137–147.
Gudas, L., Eriksson, S., Ullman, B., and Martin, D., Jr. (1981).Adv. Enzym. Regul. 19:129–137.
Bradford, M. (1976).Anal. Biochem. 72:248–254.
Berglund, O., and Eckstein, F. (1974).Methods Enzymol. 34B:253–261.
Knorre, I., Kurbatov, V., and Sanukov, V. (1976).FEBS Lett. 70:105–108.
Lewis, W., and Wright, J. (1978).J. Cell. Physiol. 78:87–98.
O'Farrell, P. (1975).J. Biol. Chem. 250:4007–4021.
Towbin, H., Staehelin, T., and Gordon, J. (1979).Proc. Natl. Acad. Sci. U.S.A. 76:4350–4354.
Johnson, D., Gautsch, J., Sportman, J., and Elder, J. (1984).Gene Anal. Tech. 1:3–8.
Standart, N., Bray, S., George, E., Hunt, T., and Ruderman, J. (1985).J. Cell. Biol. 100:1968–1976.
Kilmartin, J., Wright, B., and Milstein, C. (1982).J. Cell Biol. 93:576–582.
Levy, H., and Sober, H. (1960).Proc. Soc. Exp. Biol. Med. 103:250–252.
Michalopoulos, E., Bevilacqua, P., Stokoe, N., Powers, V., Willard, H., and Lewis, W. (1985).Hum. Genet. 70:157–162.
Srinivasan, P., Tonin, P., Wensing, E., and Lewis, W. (1987).J. Biol. Chem. (submitted).
Caras, I., Levinson, B., Fabry, M., Williams, S., and Martin, D., Jr. (1985).J. Biol. Chem. 260:7015–7022.
Feinberg, A., and Vogelstein, B. (1983).Anal. Biochem. 132:6–13.
Cory, J., Fleischer, A., and Munro, J., III. (1978).J. Biol. Chem. 253:2898–2901.
Lewis, W., Kuzik, B., and Wright, J. (1978).J. Cell. Physiol. 94:287–298.
Hopper, S. (1972).J. Biol. Chem. 247:3336–3340.
Thelander, L., Eriksson, S., and Åkerman, M. (1980).J. Biol. Chem. 255:7426–7432.
Korneluk, R., Quan, F., Lewis, W., Guise, K., Willard, H., Holmes, M., and Gravel, R. (1984).J. Biol. Chem. 259:13819–13823.
Zimmerman, C., and Gold, A. (1983).Biochemistry 22:3387–3392.
Albert, D., and Gudas, L. (1985).J. Biol. Chem. 260:679–684.
Stark, G. (1986).Cancer Surv. 5:1–23.
Alt, F., Kellems, R., Bertino, J., and Schimke, R. (1978).J. Biol. Chem. 253:1357–1370.
Steglich, C., Choi, J., and Scheffler, I. (1985). InMolecular Cell Genetics, (ed.) Gottesman, M.M. (John Wiley & Sons, New York).
Padgett, R., Wahl, G., Coleman, P., and Stark, G. (1979).J. Biol. Chem. 254:974–980.
Yang-Feng, T., Thelander, L., Lewis, W., Srinivasan, P., and Francke, U. (1986).J. Hum. Genet. 39:A174.
Tonin, P., Stallings, R., Carman, M., Bertino, J., Wright, J.A., Srinivasan, P., and Lewis, W. (1987).Cytogenet. Cell. Genet. (in press).
Engstrom, Y., Rozell, B., Hansson, H., Stemme, S., and Thelander, L. (1984).EMBO J. 3:863–867.
Eriksson, S., Gräslund, A., Skog, S., Thelander, L., and Tribukait, B. (1984).J. Biol. Chem. 259:11695–11700.
Eriksson, S., and Martin, D., Jr. (1981).J. Biol. Chem. 256:9436–9440.
Brown, N., and Eliasson, R. (1969).Eur. J. Biochem. 9:512–518.
Gräslund, A., Ehrenberg, A., and Thelander, L. (1982).J. Biol. Chem. 257:5711–5715.
Valenzuela, P., Quiroga, M., Zaldivar, J., Cleveland, D.W., Rutter, W.J., and Kirshner, M.W. (1981).Nature 289:650–655.
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Cocking, J.M., Tonin, P.N., Stokoe, N.M. et al. Gene for M1 subunit of ribonucleotide reductase is amplified in hydroxyurea-resistant hamster cells. Somat Cell Mol Genet 13, 221–233 (1987). https://doi.org/10.1007/BF01535204
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DOI: https://doi.org/10.1007/BF01535204