Antifolate-resistant Chinese hamster cells. Evidence for dihydrofolate reductase gene amplification among independently derived sublines overproducing different dihydrofolate reductases.
1980
Abstract Purification of a cDNA probe specific for dihydrofolate reductase mRNA has allowed the relative quantification of dihydrofolate reductase gene numbers and mRNA levels between a line of Chinese hamster lung fibroblasts sensitive to the antifolate drugs methotrexate and methasquin and sublines which by virtue of their overproduction of the target enzyme dihydrofolate reductase are cross-resistant to these site-specific inhibitors. Previous results have shown that resistant sublines could overproduce one of two molecular weight forms of dihydrofolate reductase, one parental-like (Mr = 21,000) and one nonparental-like (Mr = 20,000), each encoded by a different mRNA present in elevated amounts in resistant cells (Melera, P. W., Wolgemuth, D., Biedler, J. L., and Hession, C. (1980). J. Biol. Chem. 255, 319--322). The cDNA-RNA and cDNA-DNA hybridization experiments reported here confirm the elevated levels of dihydrofolate reductase-specific mRNA in resistant cells and show that the overproduction of either molecular weight form of dihydrofolate reductase is accompanied by amplification in dihydrofolate reductase gene number.
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