Abstract
Chinese hamster ovary cell lines which are resistant to an amino acid analog, beta-aspartyl hydroxamate, have been isolated and characterized. Mutants resistant to 100-150 microM beta-aspartyl hydroxamate arose from ethyl methane sulfonate-treated parental lines at frequencies of 3.4 x 10(-6) to 1.3 x 10(-7). The mutants fell into at least two genetic classes: 18% of the mutants behaved codominantly in hybrids, the others recessively. Complementation studies indicated that all the recessive mutants belonged to the same class. Mutants selected after one step of mutagenesis overproduce the enzyme asparagine synthetase constitutively with four- to sixfold increases in specific activities over the basal levels of the parental lines. beta-Aspartyl hydroxamate-resistant cell lines with up to 20-fold elevations in asparagine synthetase activity have been isolated after two steps of mutageneis. In addition, highly resistant lines have been selected by long-term growth of a dominant mutant in increasing concentrations of the drug. Resistance in the latter appears to be due not only to overproduction of asparagine synthetase but also to an alteration in the affinity of the enzyme for beta-aspartyl hydroxamate.
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