Biochemical genetics of Chinese hamster cell mutants with deviant purine metabolism: characterization of Chinese hamster cell mutants defective in phosphoribosylpyrophosphate amidotransferase and phosphoribosylglycinamide synthetase and an examination of alternatives to the first step of purine biosynthesis.
Activities of the first three enzymes in the de novo purine biosynthetic pathway have been measured in cell-free extracts of the Chinese hamster ovary cell (CHO-K1) and two purine-requiring auxotrophs of this cell. Ade-A has been found to be defective in phosphoribosylpryophosphate (PRPP) amidotransferase while Ade-C has been found to be defective in glycinamide ribonucleotide (GAR) synthetase. Neither enzyme deficiency is due to the presence of an excess of diffusible inhibitor, and mixed extracts of Ade-A and Ade-C are capable of performing both enzymatic steps in a coupled assay. Assays of GAR formyltransferase show that it is present in Ade-A and Ade-C, indicating that these cell types are defective in only one enzyme each of the early purine biosynthetic enzymes. Using the Ade-A mutant, analysis of alternatives to PRPP plus glutamine as substrates for the first step in the purine biosynthetic pathway showed that a common genetic unit must direct the synthesis for both PRPP plus glutamine and PRPP plus ammonia activities. Although ribose-5-phosphate plus ammonia can be used in cell-free extracts to perform the first step in purine biosynthesis, it is shown that this activity is apparently not used by intact CHO-K1 cells.
D000582:Amidophosphoribosyltransferase / Q000235:genetics* / Q000378:metabolism; D000588:Amines / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D008025:Ligases / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010430:Pentosyltransferases / Q000235:genetics*; D011687:Purines / Q000096:biosynthesis*; D012267:Ribosemonophosphates / Q000378:metabolism
D000588:Amines; D011687:Purines; D012267:Ribosemonophosphates; D010430:Pentosyltransferases; D000582:Amidophosphoribosyltransferase; D008025:Ligases