Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Apr;4(4):618–624. doi: 10.1128/mcb.4.4.618

Amplification of hypoxanthine-guanine phosphoribosyltransferase genes in chromosome-mediated gene transferents.

S Linder, A W Coleman, J M Eisenstadt
PMCID: PMC368767  PMID: 6717438

Abstract

Hypoxanthine-guanine phosphoribosyltransferase (HPRT) enzyme activities may be elevated in genetically unstable chromosome-mediated gene transferents selected for transfer of the HPRT gene. Increased levels of HPRT polypeptides in unstable mouse L cell gene transferents were demonstrated by two-dimensional gel electrophoresis and immunoprecipitation. No additional polypeptides were found to be overexpressed. HPRT mRNA levels were elevated 10- to 15-fold in the unstable gene transferent GT427C. Southern blot hybridization experiments showed that overexpression of HPRT correlated with a 5- to 15-fold amplification of HPRT gene sequences in two unstable cell lines. Stabilized gene transferents displayed reduced HPRT copy numbers. The amplification of HPRT gene sequences in the unstable transferent GT427C was associated with the presence of multiple minute chromosome fragments. An average of 9.6 fragments was found per metaphase, but the variation was considerable, ranging from 0 to 53. We conclude that genomic DNA sequences may be amplified in unstable chromosome-mediated gene transferents and that such amplification may be associated with the occurrence of multiple chromosomal fragments.

Full text

PDF
618

Images in this article

620Image
on p.620
621Image
on p.621
621Image
on p.621
622Image
on p.622
622Image
on p.622

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brennand J., Chinault A. C., Konecki D. S., Melton D. W., Caskey C. T. Cloned cDNA sequences of the hypoxanthine/guanine phosphoribosyltransferase gene from a mouse neuroblastoma cell line found to have amplified genomic sequences. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1950–1954. doi: 10.1073/pnas.79.6.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chasin L. A., Urlaub G. Chromosome-wide event accompanies the expression of recessive mutations in tetraploid cells. Science. 1975 Mar 21;187(4181):1091–1093. doi: 10.1126/science.1167702. [DOI] [PubMed] [Google Scholar]
  3. Coleman A. W., Maguire M. J., Coleman J. R. Mithramycin- and 4'-6-diamidino-2-phenylindole (DAPI)-DNA staining for fluorescence microspectrophotometric measurement of DNA in nuclei, plastids, and virus particles. J Histochem Cytochem. 1981 Aug;29(8):959–968. doi: 10.1177/29.8.6168681. [DOI] [PubMed] [Google Scholar]
  4. Cowell J. K. Double minutes and homogeneously staining regions: gene amplification in mammalian cells. Annu Rev Genet. 1982;16:21–59. doi: 10.1146/annurev.ge.16.120182.000321. [DOI] [PubMed] [Google Scholar]
  5. Degnen G. E., Miller I. L., Adelberg E. A., Eisenstadt J. M. Overexpression of an unstably inherited gene in cultured mouse cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3956–3959. doi: 10.1073/pnas.74.9.3956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Degnen G. E., Miller I. L., Eisenstadt J. M., Adelberg E. A. Chromosome-mediated gene transfer between closely realted strains of cultured mouse cells. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2838–2842. doi: 10.1073/pnas.73.8.2838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ghangas G. S., Milman G. Hypoxanthine phosphoribosyltransferase: two-dimensional gels from normal and Lesch-Nyhan hemolyzates. Science. 1977 Jun 3;196(4294):1119–1120. doi: 10.1126/science.870972. [DOI] [PubMed] [Google Scholar]
  8. Haber D. A., Schimke R. T. Chromosome-mediated transfer and amplification of an altered mouse dihydrofolate reductase gene. Somatic Cell Genet. 1982 Jul;8(4):499–508. doi: 10.1007/BF01538710. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Kaufman R. J., Brown P. C., Schimke R. T. Loss and stabilization of amplified dihydrofolate reductase genes in mouse sarcoma S-180 cell lines. Mol Cell Biol. 1981 Dec;1(12):1084–1093. doi: 10.1128/mcb.1.12.1084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Klobutcher L. A., Miller C. L., Ruddle F. H. Chromosome-mediated gene transfer results in two classes of unstable transformants. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3610–3614. doi: 10.1073/pnas.77.6.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klobutcher L. A., Ruddle F. H. Phenotype stabilisation and integration of transferred material in chromosome-mediated gene transfer. Nature. 1979 Aug 23;280(5724):657–660. doi: 10.1038/280657a0. [DOI] [PubMed] [Google Scholar]
  13. Linder S., Brzeski H., Ringertz N. R. Phenotypic expression in cybrids derived from teratocarcinoma cells fused with myoblast cytoplasms. Exp Cell Res. 1979 Apr;120(1):1–14. doi: 10.1016/0014-4827(79)90529-9. [DOI] [PubMed] [Google Scholar]
  14. McBride O. W., Peterson J. L. Chromosome-mediated gene transfer in mammalian cells. Annu Rev Genet. 1980;14:321–345. doi: 10.1146/annurev.ge.14.120180.001541. [DOI] [PubMed] [Google Scholar]
  15. Merril C. R., Goldman D., Ebert M. Protein variations associated with Lesch-Nyhan syndrome. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6471–6475. doi: 10.1073/pnas.78.10.6471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mose-Larsen P., Bravo R., Fey S. J., Small J. V., Celis J. E. Putative association of mitochondria with a subpopulation of intermediate-sized filaments in cultured human skin fibroblasts. Cell. 1982 Dec;31(3 Pt 2):681–692. doi: 10.1016/0092-8674(82)90323-3. [DOI] [PubMed] [Google Scholar]
  17. Musick W. D. Structural features of the phosphoribosyltransferases and their relationship to the human deficiency disorders of purine and pyrimidine metabolism. CRC Crit Rev Biochem. 1981;11(1):1–34. doi: 10.3109/10409238109108698. [DOI] [PubMed] [Google Scholar]
  18. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  19. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  20. Olsen A. S., McBride O. W., Moore D. E. Number and size of human X chromosome fragments transferred to mouse cells by chromosome-mediated gene transfer. Mol Cell Biol. 1981 May;1(5):439–448. doi: 10.1128/mcb.1.5.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ruddle F. H. A new era in mammalian gene mapping: somatic cell genetics and recombinant DNA methodologies. Nature. 1981 Nov 12;294(5837):115–120. doi: 10.1038/294115a0. [DOI] [PubMed] [Google Scholar]
  22. Sharp J. D., Capecchi N. E., Capecchi M. R. Altered enzymes in drug-resistant variants of mammalian tissue culture cells. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3145–3149. doi: 10.1073/pnas.70.11.3145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  24. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ward D. C., Reich E., Goldberg I. H. Base specificity in the interaction of polynucleotides with antibiotic drugs. Science. 1965 Sep 10;149(3689):1259–1263. doi: 10.1126/science.149.3689.1259. [DOI] [PubMed] [Google Scholar]
  26. Willecke K., Lange R., Krüger A., Reber T. Cotransfer of two linked human genes into cultured mouse cells. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1274–1278. doi: 10.1073/pnas.73.4.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wullems G. J., van der Horst J., Bootsma D. Transfer of the human genes coding for thymidine kinase and galactokinase to Chinese hamster cells and human-Chinese hamster cell hybrids. Somatic Cell Genet. 1977 May;3(3):281–293. doi: 10.1007/BF01538746. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES