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  • Title: DRB resistance in Chinese hamster and human cells: genetic and biochemical characteristics of the selection system.
    Author: Gupta RS, Siminovitch L.
    Journal: Somatic Cell Genet; 1980 Mar; 6(2):151-69. PubMed ID: 6930702.
    Abstract:
    Stable mutants resistant to the nucleoside analog 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole (DRB), which interferes with RNA synthesis, have been selected in Chinese hamster ovary (CHO) and human diploid fibroblasts. In CHO cells, upon treatment with the mutagen ethyl-methane sulfonate (EMS), a linear dose--response between the concentration of mutagen and the frequency of DrbR mutants was observed in the range of 20--300 micrograms/ml. The selection system did not show cell density or cross-feeding effects, and the optimal expression time following mutagenesis was found to be 2--3 days for CHO cells and 5--6 days for human fibroblasts. The DrbR mutation behaved codominantly in DrbR x DrbS hybrids. Addition of DRB affected nucleoside uptake to a similar extent in both wild-type and mutant cells, indicating that the drug was able to enter the mutant cells. The failure of DrbR mutants to show any cross-resistance to other toxic nucleoside analogs examined suggests that the action of DRB does not involve the initial phosphorylation step. DRB addition did not cause any marked inhibition of either RNA polymerase I or RNA polymerase II activity from both wild-type and mutant cells in vitro, indicating that its effect on RNA synthesis may be indirect.
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