Regulation of glucose 6-phosphate dehydrogenase expression in CHO-human fibroblast somatic cell hybrids.
Human--hamster somatic cell hybrids have been obtained by fusion of a CHO line (NA31) doubly deficient in hypoxanthine guanine phosphoribosyltransferase and glucose 6-phosphate dehydrogenase (G6PD) with normal G6PD(+) human fibroblasts. Analysis of NA31 extracts has revealed that, although G6PD activity is nearly absent, significant activity can be detected with 2-deoxyglucose 6-phosphate as substrate, so that the mutant and normal forms of the enzyme can both be easily detected. The cell hybrids obtained express human G6PD. The human G6PD subunits are distributed in homodimeric molecules as well as in human--hamster heterodimeric molecules. However, whereas the amount of hamster G6PD subunits present in the hybrid is similar to that in the hamster parental cells, the amount of human G6PD subunits is decreased by 3- to 10-fold when compared to the human parental cell. These results indicate that either the expression of the G6PD gene or the stability of the gene product is altered in the hybrid. By mutagenesis and selection in diamide (a substance that oxidizes intracellular glutathione), we have isolated a clone with a 3- to 5-fold increase in human G6PD activity. This derivative may have an increased rate of expression of the human G6PD structural gene.
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003958:Diamide / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000201:enzymology*; D005786:Gene Expression Regulation; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006822:Hybrid Cells / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D009154:Mutation; D014960:X Chromosome / Q000201:enzymology
D003958:Diamide; D005954:Glucosephosphate Dehydrogenase