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. 1985 Feb;82(3):780–784. doi: 10.1073/pnas.82.3.780

Adult and fetal human globin genes are expressed following chromosomal transfer into MEL cells.

T H Papayannopoulou, D Lindsley, S Kurachi, K Lewison, T Hemenway, M Melis, N P Anagnou, V Najfeld
PMCID: PMC397130  PMID: 2579380

Abstract

Somatic cell hybridization of mouse erythroleukemia (MEL) cells and HEL cells, a human erythroleukemia line that produces fetal (gamma) but fails to express adult (beta) globin, was used to test whether the expression of the two human globin genes is regulated cis or trans. An experimental approach using anti-human globin monoclonal antibodies for detection, efficient cloning, and monitoring of hybrids of interest was employed. Further characterization of hybrids used isoelectric focusing for detection of human globins and S1 nuclease mapping. In contrast to the parental HEL line, all chromosome 11-retaining HEL-MEL hybrids expressed human beta-globin, suggesting that the HEL beta-globin genes (i) are transcriptionally competent, (ii) become activated in response to a positive trans-acting element within the MEL environment, and (iii) fail to express into the HEL environment because of either the absence of a positive trans-acting element or the presence of a trans-acting inhibitor of beta-globin gene expression. In addition to beta-globin, the primary HEL-MEL hybrids co-expressed gamma-globin; however, gamma-globin expression segregated by subcloning so that secondary and tertiary clones either expressed only beta-globin or co-expressed gamma- and beta-globin. The results of subcloning can be explained by assuming that gamma-globin gene expression is controlled by a HEL cell-derived transacting element encoded by a gene not syntenic to chromosome 11 or by postulating that the HEL gamma-globin genes become randomly modified during the continuous proliferation of hybrids.

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Selected References

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

  1. Benz E. J., Jr, Murnane M. J., Tonkonow B. L., Berman B. W., Mazur E. M., Cavallesco C., Jenko T., Snyder E. L., Forget B. G., Hoffman R. Embryonic-fetal erythroid characteristics of a human leukemic cell line. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3509–3513. doi: 10.1073/pnas.77.6.3509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  3. Dean A., Ley T. J., Humphries R. K., Fordis M., Schechter A. N. Inducible transcription of five globin genes in K562 human leukemia cells. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5515–5519. doi: 10.1073/pnas.80.18.5515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Deisseroth A., Barker J., Anderson W. F., Nienhuis Hemoglobin synthesis in somatic cell hybrids: coexpression of mouse with human or chinese hamster globin genes in interspecific somatic cell hybrids of mouse erythroleukemia cells. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2682–2686. doi: 10.1073/pnas.72.7.2682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deisseroth A., Burk R., Picciano D., Anderson W. F., Nienhuis A., Minna J. Hemoglobin synthesis in somatic cell hybrids: globin gene expression in hybrids between mouse erythroleukemia and human marrow cells or fibroblasts. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1102–1106. doi: 10.1073/pnas.72.3.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deisseroth A., Hendrick D. Human alpha-globin gene expression following chromosomal dependent gene transfer into mouse erythroleukemia cells. Cell. 1978 Sep;15(1):55–63. doi: 10.1016/0092-8674(78)90082-x. [DOI] [PubMed] [Google Scholar]
  7. Deisseroth A., Velez R., Nienhuis A. W. Hemoglobin synthesis in somatic cell hybrids: independent segregation of the human alpha- and beta-globin genes. Science. 1976 Mar 26;191(4233):1262–1264. doi: 10.1126/science.943846. [DOI] [PubMed] [Google Scholar]
  8. Emerson B. M., Felsenfeld G. Specific factor conferring nuclease hypersensitivity at the 5' end of the chicken adult beta-globin gene. Proc Natl Acad Sci U S A. 1984 Jan;81(1):95–99. doi: 10.1073/pnas.81.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fordis C. M., Anagnou N. P., Dean A., Nienhuis A. W., Schechter A. N. A beta-globin gene, inactive in the K562 leukemic cell, functions normally in a heterologous expression system. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4485–4489. doi: 10.1073/pnas.81.14.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Groudine M., Kohwi-Shigematsu T., Gelinas R., Stamatoyannopoulos G., Papayannopoulou T. Human fetal to adult hemoglobin switching: changes in chromatin structure of the beta-globin gene locus. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7551–7555. doi: 10.1073/pnas.80.24.7551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ley T. J., Anagnou N. P., Pepe G., Nienhuis A. W. RNA processing errors in patients with beta-thalassemia. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4775–4779. doi: 10.1073/pnas.79.15.4775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  13. Martin P., Papayannopoulou T. HEL cells: a new human erythroleukemia cell line with spontaneous and induced globin expression. Science. 1982 Jun 11;216(4551):1233–1235. doi: 10.1126/science.6177045. [DOI] [PubMed] [Google Scholar]
  14. Meera Khan P. Enzyme electrophoresis on cellulose acetate gel: zymogram patterns in mgh-mouse and man--Chinese hamster somatic cell hybrids. Arch Biochem Biophys. 1971 Aug;145(2):470–483. doi: 10.1016/s0003-9861(71)80007-3. [DOI] [PubMed] [Google Scholar]
  15. Mueller R. F., Murray J. C., Gelinas R., Farquhar M., Papayannopoulou T. Restriction endonuclease mapping of globin genomic regions of HEL (human erythroleukemia) line. Hemoglobin. 1983;7(3):245–256. doi: 10.3109/03630268309048653. [DOI] [PubMed] [Google Scholar]
  16. Orkin S. H., Kazazian H. H., Jr, Antonarakis S. E., Goff S. C., Boehm C. D., Sexton J. P., Waber P. G., Giardina P. J. Linkage of beta-thalassaemia mutations and beta-globin gene polymorphisms with DNA polymorphisms in human beta-globin gene cluster. Nature. 1982 Apr 15;296(5858):627–631. doi: 10.1038/296627a0. [DOI] [PubMed] [Google Scholar]
  17. Pyati J., Kucherlapati R. S., Skoultchi A. I. Activation of human beta-globin genes from nonerythroid cells by fusion with murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3435–3439. doi: 10.1073/pnas.77.6.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Righetti P. G., Gianazza E., Gianni A. M., Comi P., Giglioni B., Ottolenghi S., Secchi C., Rossi-Bernardi L. Human globin chain separation by isoelectric focusing. J Biochem Biophys Methods. 1979;1(1):45–57. doi: 10.1016/0165-022x(79)90045-9. [DOI] [PubMed] [Google Scholar]
  19. Rutherford T. R., Clegg J. B., Weatherall D. J. K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature. 1979 Jul 12;280(5718):164–165. doi: 10.1038/280164a0. [DOI] [PubMed] [Google Scholar]
  20. Rutherford T., Clegg J. B., Higgs D. R., Jones R. W., Thompson J., Weatherall D. J. Embryonic erythroid differentiation in the human leukemic cell line K562. Proc Natl Acad Sci U S A. 1981 Jan;78(1):348–352. doi: 10.1073/pnas.78.1.348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Stamatoyannopoulos G., Farquhar M., Lindsley D., Brice M., Papayannopoulou T., Nute P. E. Monoclonal antibodies specific for globin chains. Blood. 1983 Mar;61(3):530–539. [PubMed] [Google Scholar]
  23. Willing M. C., Nienhuis A. W., Anderson W. F. Selective activation of human beta-but not gamma-globin gene in human fibroblast x mouse erythroleukaemia cell hybrids. Nature. 1979 Feb 15;277(5697):534–538. doi: 10.1038/277534a0. [DOI] [PubMed] [Google Scholar]
  24. Wright S., deBoer E., Grosveld F. G., Flavell R. A. Regulated expression of the human beta-globin gene family in murine erythroleukaemia cells. Nature. 1983 Sep 22;305(5932):333–336. doi: 10.1038/305333a0. [DOI] [PubMed] [Google Scholar]

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