Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Apr;79(7):2281–2285. doi: 10.1073/pnas.79.7.2281

More than one-third of the discernible mouse polypeptides are not expressed in a Chinese hamster-mouse embryo fibroblast hybrid that retains all mouse chromosomes.

R Bravo, R Schafer, K Willecke, H MacDonald-Bravo, S J Fey, J E Celis
PMCID: PMC346176  PMID: 6954539

Abstract

Gene expression at the level of polypeptide synthesis has been investigated in a somatic cell hybrid (20 BW-4) isolated after fusion of spontaneously transformed, tumorigenic Chinese hamster lung fibroblasts with mouse embryo fibroblasts. This hybrid exhibited suppression of tumorigenicity and retained--in addition to the parental Chinese hamster genome--copies of all mouse chromosomes as demonstrated by direct karyotype analysis and confirmed for 18 different mouse chromosomes by analysis of 18 different mouse isozymes. Two-dimensional gel electrophoresis of [35S]methionine-labeled polypeptides from hybrid 20 BW-4 showed that the overall polypeptide pattern corresponded to that of the hamster parent. All polypeptides detected in the hamster parental cells were also expressed in the hybrid although some of them were expressed in altered amounts. Of approximately 1200 labeled polypeptides revealed in the parental cells, 115 mouse polypeptides could be clearly distinguished from the hamster polypeptides due to their different electrophoretic mobilities. Forty-two of these (i.e., 37%) were not expressed in the hybrid 20 BW-4. These observations were confirmed by analysis of another independently isolated hybrid (2W 23) of the same parental cells that also exhibited suppression of malignancy and that retained copies of all mouse chromosomes except no. 5. The results suggest that the genome of the tumorigenic cell after hybridization can suppress expression of more than one-third of the normal parental genome. The suppressed mouse genetic information is probably located on many, if not all, different mouse chromosomes. Even if the level of genetic suppression is high the mouse genome is able to reduce the tumorigenicity of the hamster parental cell.

Full text

PDF
2281

Images in this article

Selected References

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

  1. Bravo R., Bellatin J., Celis J. E. [35S]-methionine labelled polypeptides from HELA cells. Coordinates and percentage of some major polypeptides. Cell Biol Int Rep. 1981 Jan;5(1):93–96. doi: 10.1016/0309-1651(81)90162-4. [DOI] [PubMed] [Google Scholar]
  2. Bravo R., Celis J. E. A search for differential polypeptide synthesis throughout the cell cycle of HeLa cells. J Cell Biol. 1980 Mar;84(3):795–802. doi: 10.1083/jcb.84.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bravo R., Fey S. J., Celis J. E. Gene expression in murine hybrids exhibiting different morphologies and tumorigenic properties. Carcinogenesis. 1981;2(8):769–782. doi: 10.1093/carcin/2.8.769. [DOI] [PubMed] [Google Scholar]
  4. Bravo R., Fey S. J., Small J. V., Larsen P. M., Celis J. E. Coexistence of three major isoactins in a single sarcoma 180 cell. Cell. 1981 Jul;25(1):195–202. doi: 10.1016/0092-8674(81)90244-0. [DOI] [PubMed] [Google Scholar]
  5. Davies P. J., Willecke K. Segregation of human hypoxanthine phosphoribosyltransferase activity from somatic cell hybrids isolated from fusion of mouse gene transfer cells with Chinese hamster cells. Mol Gen Genet. 1977 Jul 20;154(2):191–197. doi: 10.1007/BF00330836. [DOI] [PubMed] [Google Scholar]
  6. Fey S. J., Bravo R., Larsen P. M., Bellatin J., Celis J. E. [35S]-methionine labelled polypeptides from secondary mouse kidney fibroblasts: coordinates and one dimensional peptide maps of some major polypeptides. Cell Biol Int Rep. 1981 May;5(5):491–500. doi: 10.1016/0309-1651(81)90176-4. [DOI] [PubMed] [Google Scholar]
  7. Francke U., Oliver N. Quantitative analysis of high-resolution trypsin-giemsa bands on human prometaphase chromosomes. Hum Genet. 1978 Dec 18;45(2):137–165. doi: 10.1007/BF00286957. [DOI] [PubMed] [Google Scholar]
  8. Kozak C. A., Lawrence J. B., Ruddle F. H. A sequential staining technique for the chromosomal analysis of the interspecific mouse/hamster and mouse/human somatic cell hybrids. Exp Cell Res. 1977 Mar 1;105(1):109–117. doi: 10.1016/0014-4827(77)90156-2. [DOI] [PubMed] [Google Scholar]
  9. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Russell W. C., Newman C., Williamson D. H. A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature. 1975 Feb 6;253(5491):461–462. doi: 10.1038/253461a0. [DOI] [PubMed] [Google Scholar]
  13. SZYBALSKA E. H., SZYBALSKI W. Genetics of human cess line. IV. DNA-mediated heritable transformation of a biochemical trait. Proc Natl Acad Sci U S A. 1962 Dec 15;48:2026–2034. doi: 10.1073/pnas.48.12.2026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schäfer R., Doehmer J., Drüge P. M., Rademacher I., Willecke K. Genetic analysis of transformed and malignant phenotypes in somatic cell hybrids between tumorigenic Chinese hamster cells and diploid mouse fibroblasts. Cancer Res. 1981 Mar;41(3):1214–1221. [PubMed] [Google Scholar]
  15. Taggart R. T., Tetri P., Francke U. Assignment of the gene for NADH diaphorase Dia-1 to Mouse chromosome 15. Somatic Cell Genet. 1980 Nov;6(6):769–776. doi: 10.1007/BF01538975. [DOI] [PubMed] [Google Scholar]
  16. Willecke K., Klomfass M., Mierau R., Döhmer J. Intraspecies transfer via total cellular DNA of the gene for hypoxanthine phosphoribosyltransferase into cultured mouse cells. Mol Gen Genet. 1979 Feb 26;170(2):179–185. doi: 10.1007/BF00337794. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES