Isolation and characterization of 5-fluorouracil-resistant mutants of Chinese hamster ovary cells deficient in the activities of orotate phosphoribosyltransferase and orotidine 5?-monophosphate decarboxylase
- 1 January 1980
- journal article
- research article
- Published by Springer Nature in Somatic Cell and Molecular Genetics
- Vol. 6 (1) , 101-114
- https://doi.org/10.1007/bf01538699
Abstract
A rapid, simple method for isolation of large numbers of Chinese hamster ovary cell (CHO-K1) mutants deficient in the final two enzymes of UMP biosynthesis, orotate phosphoribosyltransferase (EC 2.4.2. 10), and OMP decarboxylase (EC 4.1.1.23) is described. The method takes advantage of the fact that CHO-K1 cells require orotate phosphoribosyltransferase to activate the pyrimidine analog 5-fluorouracil to its active form; hence, mutants lacking this enzyme activity can be isolated simply by selection for cells resistant to 5-fluorouracil. The relevance of these observations to the metabolism of this cancer chemotheraputic agent, to the study of the structure of the protein(s) involved in catalyzing the last two steps of UMP biosynthesis, to the study of the structure of the gene(s) coding for this protein, and to analysis of the human genetic disease orotic aciduria is discussed.This publication has 41 references indexed in Scilit:
- Biochemical genetic analysis of pyrimidine biosynthesis in mammalian cells. II. Isolation and characterization of a mutant of chinese hamster ovary cells with defective dihydroorotate dehydrogenase (E.C. 1.3.3.1) activityJournal of Cellular Physiology, 1979
- Mutagenesis of Chinese hamster cells is facilitated by thymidine and deoxycytidineNature, 1978
- Subunit structure of the orotate phosphoribosyltransferase-orotidylate decarboxylase complex from human erythrocytesBiochemistry, 1977
- Stable mutants of mammalian cells that overproduce the first three enzymes of pyrimidine nucleotide biosynthesisCell, 1976
- A replica plating method for CHO cells using nylon clothCytogenetic and Genome Research, 1975
- Uridylic acid synthesis in Ehrlich ascites carcinoma. Properties, subcellular distribution, and nature of enzyme complexes of the six biosynthetic enzymesBiochemistry, 1973
- Resistance of Mammalian Tumour Cells toward Pyrimidine AnaloguesOncology, 1973
- Alteration of quaternary structural behaviour of an hepatic orotate phosphoribosyltransferase-orotidine-5′-phosphate decarboxylase complex in rats following allopurinol therapyBiochemical Pharmacology, 1972
- Regulation of pyrimidine and arginine biosynthesis in mammalsAdvances in Enzyme Regulation, 1971
- Genetics of somatic mammalian cells. IX. Quantitation of mutagenesis by physical and chemical agentsJournal of Cellular Physiology, 1969