Abstract
Mutants unable to perform de novo biosynthesis of purines have been isolated from cultures of mutagen-treated Chinese hamster ovary cells using bromodeoxyuridine selection techniques. Accumulation of C14-labeled formylglycinamide ribotide by suspension cultures of mutant cells incubated with glycine-C14 suggested that the defect leading to auxotrophy most probably involves the gene coding for formylglycinamide amidotransferase, (E.C. 6.3, 5.3), the fourth enzyme in the de novo purine biosynthetic pathway. Direct assay of formylglycinamide amidotransferase activity in cell-free extracts prepared from mutant and parental cells has demonstrated the absence of amidotransferase activity in mutant derived extracts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Buchanan, J. M. (1973). The amidotransferases. In Meister, A. (ed.), Advances in Enzymology, Vol. 39, John Wiley and Sons, New York.
Flaks, J., and Lukens, L. (1963). The enzymes of purine nucleotide synthesis de novo. In Colowick, S. P., and Kaplan, N. O. (eds.), Methods in Enzymology, Vol. VI, Academic Press, New York.
Goldthwait, D. A., Peabody, R. A., and Greenberg, G. R. (1956). On the occurrence of glycinamide ribotide and its formyl derivative. J. Biol. Chem. 221555.
Henderson, J. F. (1962). Feedback inhibition of purine biosynthesis. J. Biol. Chem. 2372631.
Lukens, L., and Flaks, J. (1963). Intermediates in purine nucleotide synthesis. In Colowick, S. P., and Kaplan, N. O. (eds.), Methods in Enzymology, Vol. VI, Academic Press, New York.
Martin, D. W., Jr., and Owen, N.T. (1972). Repression and derepression of purine biosynthesis in mammalian hepatoma cells in culture. J. Biol. Chem. 2475477.
Paul, J. (1972). Cell and Tissue Culture, 4th ed. Williams and Wilkins, Baltimore.
Taylor, M. W., Souhrada, M., and McCall, J. (1971). New class of purine mutants of Chinese hamster ovary cells. Science 172162.
Wyngaarden, J. B. (1972). Glutamine phosphoribosylpyrophosphate amidotransferase. In Horecker, B. L., and Stadtman, E. R. (eds.), Current Topics in Cellular Regulation, Vol. 5, Academic Press, New York.
Author information
Authors and Affiliations
Additional information
This work was supported by Public Health Service Grant No. GM 18924.
Rights and permissions
About this article
Cite this article
Feldman, R.I., Taylor, M.W. Purine mutants of mammalian cell lines. I. Accumulation of formylglycinamide ribotide by purine mutants of Chinese hamster ovary cells. Biochem Genet 12, 393–405 (1974). https://doi.org/10.1007/BF00486644
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00486644