Isolation and Characterization of Mutants of Human Diploid Fibroblasts Resistant to Diphtheria Toxin

Stable mutants highly resistant to the protein synthesis inhibitor diphtheria toxin (dipr) have been selected in human diploid fibroblast cells at a frequency of 1-8 × 10-6. Treatment of cells with mutagens, (e.g., ethylmethanesulfonate, nitrosoguanidine, and ICR-170), increased the frequencies of d...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1978-07, Vol.75 (7), p.3337-3340
Main Authors: Gupta, Radhey S., Siminovitch, Louis
Format: Article
Language:English
Subjects:
Cell Line
Cells - drug effects
CHO cells
Cultured cells
Diphtheria
Diphtheria Toxin - pharmacology
Diploidy
Drug Resistance
Fibroblasts
Genetic mutation
Humans
KB cells
Mutagens
Mutation
Peptide Elongation Factors
Protein Biosynthesis
Protein synthesis
Somatic cells
Toxins
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Summary:Stable mutants highly resistant to the protein synthesis inhibitor diphtheria toxin (dipr) have been selected in human diploid fibroblast cells at a frequency of 1-8 × 10-6. Treatment of cells with mutagens, (e.g., ethylmethanesulfonate, nitrosoguanidine, and ICR-170), increased the frequencies of diprmutants by 50- to 500-fold in different experiments, and the optimal expression time for diprmutation was about 5 days. All mutants examined thus far have bred true, and no effects of cell density or cross feeding have been observed on the selection. Fluctuation analysis showed that the diprmutation occurs in these fibroblasts at the rate of 5-6 × 10-7mutations per cell per generation. Protein synthesis in mutant extracts was resistant to diphtheria toxin, indicating that the diprlesion in such mutants lies in the protein synthesis machinery. The characteristics of the diprmarker should make this system particularly useful for studies of quantitative mutagenesis in human diploid cells.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.75.7.3337