Abstract
The amino acid analog albizziin was used to isolate Chinese hamster ovary cell lines which overproduce asparagine synthetase. Mutants selected in a single step after ethyl methane sulfonate mutagenesis were approximately 10-fold more resistant to the drug than the parental lines and expressed 8- to 17-fold elevations in enzyme activity. The karyotypes of these lines show alterations such as breaks and translocations affecting the long arm of chromosome 1. Cell lines isolated in several steps by growth in progressively increasing concentrations of albizziin were more resistant to the drug and exhibited up to 300-fold enhancement of asparagine synthetase activity. The multistep albizziin-resistant cell lines usually had expanded chromosomal regions which stained somewhat homogeneously, often on the long arm of chromosome 1. These results suggest that resistance to albizziin in the multistep lines may be due to gene amplification.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alt F. W., Kellems R. E., Bertino J. R., Schimke R. T. Selective multiplication of dihydrofolate reductase genes in methotrexate-resistant variants of cultured murine cells. J Biol Chem. 1978 Mar 10;253(5):1357–1370. [PubMed] [Google Scholar]
- Andrulis I. L., Siminovitch L. DNA-mediated gene transfer of beta-aspartylhydroxamate resistance into Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5724–5728. doi: 10.1073/pnas.78.9.5724. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Andrulis I. L., Siminovitch L. Isolation and characterization of Chinese hamster ovary cell mutants resistant to the amino acid analog beta-aspartyl hydroxamate. Somatic Cell Genet. 1982 Jul;8(4):533–545. doi: 10.1007/BF01538713. [DOI] [PubMed] [Google Scholar]
- Biedler J. L., Spengler B. A. Metaphase chromosome anomaly: association with drug resistance and cell-specific products. Science. 1976 Jan 16;191(4223):185–187. doi: 10.1126/science.942798. [DOI] [PubMed] [Google Scholar]
- Flintoff W. F., Davidson S. V., Siminovitch L. Isolation and partial characterization of three methotrexate-resistant phenotypes from Chinese hamster ovary cells. Somatic Cell Genet. 1976 May;2(3):245–261. doi: 10.1007/BF01538963. [DOI] [PubMed] [Google Scholar]
- Flintoff W. F., Weber M. K., Nagainis C. R., Essani A. K., Robertson D., Salser W. Overproduction of dihydrofolate reductase and gene amplification in methotrexate-resistant Chinese hamster ovary cells. Mol Cell Biol. 1982 Mar;2(3):275–285. doi: 10.1128/mcb.2.3.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gantt J. S., Chiang C. S., Hatfield G. W., Arfin S. M. Chinese hamster ovary cells resistant to beta-aspartylhydroxamate contain increased levels of asparagine synthetase. J Biol Chem. 1980 May 25;255(10):4808–4813. [PubMed] [Google Scholar]
- Horowitz B., Meister A. Glutamine-dependent asparagine synthetase from leukemia cells. Chloride dependence, mechanism of action, and inhibition. J Biol Chem. 1972 Oct 25;247(20):6708–6719. [PubMed] [Google Scholar]
- Kaufman R. J., Brown P. C., Schimke R. T. Amplified dihydrofolate reductase genes in unstably methotrexate-resistant cells are associated with double minute chromosomes. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5669–5673. doi: 10.1073/pnas.76.11.5669. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaufman R. J., Schimke R. T. Amplification and loss of dihydrofolate reductase genes in a Chinese hamster ovary cell line. Mol Cell Biol. 1981 Dec;1(12):1069–1076. doi: 10.1128/mcb.1.12.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kellems R. E., Morhenn V. B., Pfendt E. A., Alt F. W., Schimke R. T. Polyoma virus and cyclic AMP-mediated control of dihydrofolate reductase mRNA abundance in methotrexate-resistant mouse fibroblasts. J Biol Chem. 1979 Jan 25;254(2):309–318. [PubMed] [Google Scholar]
- Kempe T. D., Swyryd E. A., Bruist M., Stark G. R. Stable mutants of mammalian cells that overproduce the first three enzymes of pyrimidine nucleotide biosynthesis. Cell. 1976 Dec;9(4 Pt 1):541–550. doi: 10.1016/0092-8674(76)90036-2. [DOI] [PubMed] [Google Scholar]
- Kopnin B. P. Specific karyotypic alterations in colchicine-resistant cells. Cytogenet Cell Genet. 1981;30(1):11–14. doi: 10.1159/000131582. [DOI] [PubMed] [Google Scholar]
- McBurney M. W., Whitmore G. F. Isolation and biochemical characterization of folate deficient mutants of Chinese hamster cells. Cell. 1974 Jul;2(3):173–182. doi: 10.1016/0092-8674(74)90091-9. [DOI] [PubMed] [Google Scholar]
- Nunberg J. H., Kaufman R. J., Schimke R. T., Urlaub G., Chasin L. A. Amplified dihydrofolate reductase genes are localized to a homogeneously staining region of a single chromosome in a methotrexate-resistant Chinese hamster ovary cell line. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5553–5556. doi: 10.1073/pnas.75.11.5553. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Padgett R. A., Wahl G. M., Coleman P. F., Stark G. R. N-(Phosphonacetyl)-L-aspartate-resistant hamster cells overaccumulate a single mRNA coding for the multifunctional protein that catalyzes the first steps of UMP synthesis. J Biol Chem. 1979 Feb 10;254(3):974–980. [PubMed] [Google Scholar]
- Pinkus L. M. Glutamine binding sites. Methods Enzymol. 1977;46:414–427. doi: 10.1016/s0076-6879(77)46049-x. [DOI] [PubMed] [Google Scholar]
- Schroeder D. D., Allison A. J., Buchanan J. M. Biosynthesis of the purines. XXXII. Effect of albizziin and other reagents on the activity of formylglycinamide ribonucleotide amidotransferase. J Biol Chem. 1969 Nov 10;244(21):5856–5865. [PubMed] [Google Scholar]
- Stanners C. P., Eliceiri G. L., Green H. Two types of ribosome in mouse-hamster hybrid cells. Nat New Biol. 1971 Mar 10;230(10):52–54. doi: 10.1038/newbio230052a0. [DOI] [PubMed] [Google Scholar]
- Wahl G. M., Padgett R. A., Stark G. R. Gene amplification causes overproduction of the first three enzymes of UMP synthesis in N-(phosphonacetyl)-L-aspartate-resistant hamster cells. J Biol Chem. 1979 Sep 10;254(17):8679–8689. [PubMed] [Google Scholar]
- Wahl G. M., Vitto L., Padgett R. A., Stark G. R. Single-copy and amplified CAD genes in Syrian hamster chromosomes localized by a highly sensitive method for in situ hybridization. Mol Cell Biol. 1982 Mar;2(3):308–319. doi: 10.1128/mcb.2.3.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Worton R. G., Duff C. Karyotyping. Methods Enzymol. 1979;58:322–344. doi: 10.1016/s0076-6879(79)58148-8. [DOI] [PubMed] [Google Scholar]
- Worton R., Duff C., Flintoff W. Microcell-mediated cotransfer of genes specifying methotrexate resistance, emetine sensitivity, and chromate sensitivity with Chinese hamster chromosome 2. Mol Cell Biol. 1981 Apr;1(4):330–335. doi: 10.1128/mcb.1.4.330. [DOI] [PMC free article] [PubMed] [Google Scholar]