Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1990 Jul;10(7):3591–3595. doi: 10.1128/mcb.10.7.3591

Differential expression of alpha- and beta-globin genes in erythroleukemic cell lines.

N Beru 1, P B Maples 1, O Hermine 1, E Goldwasser 1
PMCID: PMC360795  PMID: 2355917

Abstract

The IW32, NN10, and IW201 cell lines are erythroleukemic cell lines isolated from the spleens of mice infected with the Friend virus. IW32 and NN10 cells can be induced toward erythroid differentiation and hemoglobin synthesis by hemin or butyrate. Both cell lines contain some mature alpha- and beta-globin mRNA before induction, and addition of the inducers greatly increases the amount of globin message. Unlike IW32 and NN10 cells, IW201 cells are only partially inducible. Uninduced 201 cells contain a small amount of alpha-globin mRNA but no detectable beta-globin message. After induction, the cells contain markedly increased amounts of alpha-globin mRNA but still do not express the beta-globin gene. Southern blot analysis with 10 restriction enzymes shows that the restriction map of the beta-globin gene in IW201 cells is indistinguishable from that in IW32 and NN10 cells.

Full text

PDF
3591

Images in this article

Selected References

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

  1. Beru N., Goldwasser E. The regulation of heme biosynthesis during erythropoietin-induced erythroid differentiation. J Biol Chem. 1985 Aug 5;260(16):9251–9257. [PubMed] [Google Scholar]
  2. Beru N., McDonald J., Goldwasser E. Studies of the constitutive expression of the mouse erythropoietin gene. Ann N Y Acad Sci. 1989;554:29–35. doi: 10.1111/j.1749-6632.1989.tb22406.x. [DOI] [PubMed] [Google Scholar]
  3. Beru N., McDonald J., Lacombe C., Goldwasser E. Expression of the erythropoietin gene. Mol Cell Biol. 1986 Jul;6(7):2571–2575. doi: 10.1128/mcb.6.7.2571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beru N., Sahr K., Goldwasser E. Inhibition of heme synthesis in bone marrow cells by succinylacetone: effect on globin synthesis. J Cell Biochem. 1983;21(2):93–105. doi: 10.1002/jcb.240210201. [DOI] [PubMed] [Google Scholar]
  5. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cai S. P., Zhang J. Z., Doherty M., Kan Y. W. A new TATA box mutation detected at prenatal diagnosis for beta-thalassemia. Am J Hum Genet. 1989 Jul;45(1):112–114. [PMC free article] [PubMed] [Google Scholar]
  7. Charnay P., Treisman R., Mellon P., Chao M., Axel R., Maniatis T. Differences in human alpha- and beta-globin gene expression in mouse erythroleukemia cells: the role of intragenic sequences. Cell. 1984 Aug;38(1):251–263. doi: 10.1016/0092-8674(84)90547-6. [DOI] [PubMed] [Google Scholar]
  8. Chehab F. F., Winterhalter K. H., Kan Y. W. Characterization of a spontaneous mutation in beta-thalassemia associated with advanced paternal age. Blood. 1989 Aug 1;74(2):852–854. [PubMed] [Google Scholar]
  9. 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]
  10. Choppin J., Casadevall N., Lacombe C., Wendling F., Goldwasser E., Berger R., Tambourin P., Varet B. Production of erythropoietin by cloned malignant murine erythroid cells. Exp Hematol. 1985 Aug;13(7):610–615. [PubMed] [Google Scholar]
  11. Choppin J., Lacombe C., Casadevall N., Muller O., Tambourin P., Varet B. Characterization of erythropoietin produced by IW32 murine erythroleukemia cells. Blood. 1984 Aug;64(2):341–347. [PubMed] [Google Scholar]
  12. Curtin P. T., Kan Y. W. The inactive beta globin gene on a gamma delta beta thalassemia chromosome has a normal structure and functions normally in vitro. Blood. 1988 Mar;71(3):766–770. [PubMed] [Google Scholar]
  13. Curtin P., Pirastu M., Kan Y. W., Gobert-Jones J. A., Stephens A. D., Lehmann H. A distant gene deletion affects beta-globin gene function in an atypical gamma delta beta-thalassemia. J Clin Invest. 1985 Oct;76(4):1554–1558. doi: 10.1172/JCI112136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Donovan-Peluso M., Young K., Dobkin C., Bank A. Erythroleukemia (K562) cells contain a functional beta-globin gene. Mol Cell Biol. 1984 Nov;4(11):2553–2555. doi: 10.1128/mcb.4.11.2553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Forrester W. C., Takegawa S., Papayannopoulou T., Stamatoyannopoulos G., Groudine M. Evidence for a locus activation region: the formation of developmentally stable hypersensitive sites in globin-expressing hybrids. Nucleic Acids Res. 1987 Dec 23;15(24):10159–10177. doi: 10.1093/nar/15.24.10159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fuchs O., Ponka P., Borova J., Neuwirt J., Travnicek M. Effect of heme on globin messenger RNA synthesis in spleen erythroid cells. J Supramol Struct Cell Biochem. 1981;15(1):73–81. doi: 10.1002/jsscb.1981.380150108. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  20. Profous-Juchelka H. R., Reuben R. C., Marks P. A., Rifkind R. A. Transcriptional and post-transcriptional regulation of globin gene accumulation in murine erythroleukemia cells. Mol Cell Biol. 1983 Feb;3(2):229–232. doi: 10.1128/mcb.3.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  22. Salditt-Georgieff M., Sheffery M., Krauter K., Darnell J. E., Jr, Rifkind R., Marks P. A. Induced transcription of the mouse beta-globin transcription unit in erythroleukemia cells. Time-course of induction and of changes in chromatin structure. J Mol Biol. 1984 Feb 5;172(4):437–450. doi: 10.1016/s0022-2836(84)80016-9. [DOI] [PubMed] [Google Scholar]
  23. Semenza G. L., Delgrosso K., Poncz M., Malladi P., Schwartz E., Surrey S. The silent carrier allele: beta thalassemia without a mutation in the beta-globin gene or its immediate flanking regions. Cell. 1984 Nov;39(1):123–128. doi: 10.1016/0092-8674(84)90197-1. [DOI] [PubMed] [Google Scholar]
  24. Sheffery M., Rifkind R. A., Marks P. A. Murine erythroleukemia cell differentiation: DNase I hypersensitivity and DNA methylation near the globin genes. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1180–1184. doi: 10.1073/pnas.79.4.1180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Tambourin P., Casadevall N., Choppin J., Lacombe C., Heard J. M., Fichelson S., Wendling F., Varet B. Production of erythropoietin-like activity by a murine erythroleukemia cell line. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6269–6273. doi: 10.1073/pnas.80.20.6269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tsai S. F., Martin D. I., Zon L. I., D'Andrea A. D., Wong G. G., Orkin S. H. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells. Nature. 1989 Jun 8;339(6224):446–451. doi: 10.1038/339446a0. [DOI] [PubMed] [Google Scholar]
  28. Tuan D., Solomon W., Li Q., London I. M. The "beta-like-globin" gene domain in human erythroid cells. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6384–6388. doi: 10.1073/pnas.82.19.6384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wright S., Rosenthal A., Flavell R., Grosveld F. DNA sequences required for regulated expression of beta-globin genes in murine erythroleukemia cells. Cell. 1984 Aug;38(1):265–273. doi: 10.1016/0092-8674(84)90548-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES