Selective amplification of polymorphic dihydrofolate reductase gene loci in Chinese hamster lung cells

Abstract

A series of antifolate-resistant Chinese hamster lung sublines that overproduce either a Mr 20,000 or a Mr 21,000 class of dihydrofolate reductase (DHFR; tetrahydrofolate dehydrogenase; 5,6,7,8-tetrahydrofolate:NADP+ oxidoreductase, EC 1.5.1.3), known to contain amplified DHFR genes, has been analyzed by DNA and RNA transfer techniques. The results suggest that the Mr 20,000 and Mr 21,000 DHFRs are encoded by at least two polymorphic DHFR genes, both of which are expressed in drug-sensitive parental cells. In drug-resistant sublines only one of the two DHFR gene types is amplified, thus accounting for the overproduction of one or the other molecular weight class of DHFR. In addition to the known differences between the DHFRs whose overproduction they direct, these allelic genes differ in restriction endonuclease profiles and in the relative abundances of their multiple mRNA transcripts.