Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 Dec 9;16(23):11027–11045. doi: 10.1093/nar/16.23.11027

Erythroid specific activation of the Xenopus laevis adult alpha-globin promoter in transient heterokaryons.

J Stalder 1
PMCID: PMC338994  PMID: 3205739

Abstract

Insertion of 1.5 kb of the 5' flanking region of the adult alpha-globin gene of X. laevis in front of the CAT structural gene promotes synthesis of CAT in transiently transfected X. laevis kidney cells. Fusion of transiently transfected kidney cells with erythroblasts isolated from anaemic frogs stimulates CAT expression 3-4 fold in the resulting transient heterokaryons. The stimulation is specific for the alpha-globin promoter and is obtained after fusion with erythroid cells but not with hepatocytes or kidney cells. Stably transfected kidney cells express drastically reduced CAT activity as compared with transiently transfected cells. Nevertheless, fusion of stably transfected kidney cells with erythroblasts leads to a 10-17 fold stimulation of CAT expression. The experiments suggest that erythroid specific transacting factors stimulate expression of CAT controlled by the adult alpha-globin promoter.

Full text

PDF
11027

Images in this article

11033Image
on p.11033
11035Image
on p.11035
11038Image
on p.11038
11039Image
on p.11039
11040Image
on p.11040
11041Image
on p.11041

Selected References

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

  1. Banerji J., Olson L., Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983 Jul;33(3):729–740. doi: 10.1016/0092-8674(83)90015-6. [DOI] [PubMed] [Google Scholar]
  2. Baron M. H., Maniatis T. Rapid reprogramming of globin gene expression in transient heterokaryons. Cell. 1986 Aug 15;46(4):591–602. doi: 10.1016/0092-8674(86)90885-8. [DOI] [PubMed] [Google Scholar]
  3. Blau H. M., Chiu C. P., Webster C. Cytoplasmic activation of human nuclear genes in stable heterocaryons. Cell. 1983 Apr;32(4):1171–1180. doi: 10.1016/0092-8674(83)90300-8. [DOI] [PubMed] [Google Scholar]
  4. Blau H. M., Pavlath G. K., Hardeman E. C., Chiu C. P., Silberstein L., Webster S. G., Miller S. C., Webster C. Plasticity of the differentiated state. Science. 1985 Nov 15;230(4727):758–766. doi: 10.1126/science.2414846. [DOI] [PubMed] [Google Scholar]
  5. Brown J. L., Ingram V. M. Structural studies on chick embryonic hemoglobins. J Biol Chem. 1974 Jun 25;249(12):3960–3972. [PubMed] [Google Scholar]
  6. 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]
  7. Choi O. R., Engel J. D. A 3' enhancer is required for temporal and tissue-specific transcriptional activation of the chicken adult beta-globin gene. Nature. 1986 Oct 23;323(6090):731–734. doi: 10.1038/323731a0. [DOI] [PubMed] [Google Scholar]
  8. Cohen R. B., Sheffery M., Kim C. G. Partial purification of a nuclear protein that binds to the CCAAT box of the mouse alpha 1-globin gene. Mol Cell Biol. 1986 Mar;6(3):821–832. doi: 10.1128/mcb.6.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crabb D. W., Dixon J. E. A method for increasing the sensitivity of chloramphenicol acetyltransferase assays in extracts of transfected cultured cells. Anal Biochem. 1987 May 15;163(1):88–92. doi: 10.1016/0003-2697(87)90096-0. [DOI] [PubMed] [Google Scholar]
  10. Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gritz L., Davies J. Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene. 1983 Nov;25(2-3):179–188. doi: 10.1016/0378-1119(83)90223-8. [DOI] [PubMed] [Google Scholar]
  13. Harlow E., Crawford L. V., Pim D. C., Williamson N. M. Monoclonal antibodies specific for simian virus 40 tumor antigens. J Virol. 1981 Sep;39(3):861–869. doi: 10.1128/jvi.39.3.861-869.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hesse J. E., Nickol J. M., Lieber M. R., Felsenfeld G. Regulated gene expression in transfected primary chicken erythrocytes. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4312–4316. doi: 10.1073/pnas.83.12.4312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hosbach H. A., Wyler T., Weber R. The Xenopus laevis globin gene family: chromosomal arrangement and gene structure. Cell. 1983 Jan;32(1):45–53. doi: 10.1016/0092-8674(83)90495-6. [DOI] [PubMed] [Google Scholar]
  16. Humphries R. K., Ley T., Turner P., Moulton A. D., Nienhuis A. W. Differences in human alpha-, beta- and delta-globin gene expression in monkey kidney cells. Cell. 1982 Aug;30(1):173–183. doi: 10.1016/0092-8674(82)90023-x. [DOI] [PubMed] [Google Scholar]
  17. Jantzen H. M., Strähle U., Gloss B., Stewart F., Schmid W., Boshart M., Miksicek R., Schütz G. Cooperativity of glucocorticoid response elements located far upstream of the tyrosine aminotransferase gene. Cell. 1987 Apr 10;49(1):29–38. doi: 10.1016/0092-8674(87)90752-5. [DOI] [PubMed] [Google Scholar]
  18. Jeffreys A. J., Wilson V., Wood D., Simons J. P., Kay R. M., Williams J. G. Linkage of adult alpha- and beta-globin genes in X. laevis and gene duplication by tetraploidization. Cell. 1980 Sep;21(2):555–564. doi: 10.1016/0092-8674(80)90493-6. [DOI] [PubMed] [Google Scholar]
  19. Karlsson S., Nienhuis A. W. Developmental regulation of human globin genes. Annu Rev Biochem. 1985;54:1071–1108. doi: 10.1146/annurev.bi.54.070185.005231. [DOI] [PubMed] [Google Scholar]
  20. Lufkin T., Bancroft C. Identification by cell fusion of gene sequences that interact with positive trans-acting factors. Science. 1987 Jul 17;237(4812):283–286. doi: 10.1126/science.3474782. [DOI] [PubMed] [Google Scholar]
  21. Maeda H., Kitamura D., Kudo A., Araki K., Watanabe T. Trans-acting nuclear protein responsible for induction of rearranged human immunoglobulin heavy chain gene. Cell. 1986 Apr 11;45(1):25–33. doi: 10.1016/0092-8674(86)90534-9. [DOI] [PubMed] [Google Scholar]
  22. Maniatis T., Goodbourn S., Fischer J. A. Regulation of inducible and tissue-specific gene expression. Science. 1987 Jun 5;236(4806):1237–1245. doi: 10.1126/science.3296191. [DOI] [PubMed] [Google Scholar]
  23. Marks P. A., Rifkind R. A. Protein synthesis: its control in erythropoiesis. Science. 1972 Mar 3;175(4025):955–961. doi: 10.1126/science.175.4025.955. [DOI] [PubMed] [Google Scholar]
  24. Mellon P., Parker V., Gluzman Y., Maniatis T. Identification of DNA sequences required for transcription of the human alpha 1-globin gene in a new SV40 host-vector system. Cell. 1981 Dec;27(2 Pt 1):279–288. doi: 10.1016/0092-8674(81)90411-6. [DOI] [PubMed] [Google Scholar]
  25. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rutherford T., Nienhuis A. W. Human globin gene promoter sequences are sufficient for specific expression of a hybrid gene transfected into tissue culture cells. Mol Cell Biol. 1987 Jan;7(1):398–402. doi: 10.1128/mcb.7.1.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Stalder J., Meyerhof W., Wirthmüller U., Gruber A., Wyler T., Knöchel W., Weber R. Conserved sequences and cell-specific DNase I hypersensitive sites upstream from the co-ordinately expressed alpha I- and alpha II-globin genes of Xenopus laevis. J Mol Biol. 1986 Mar 20;188(2):119–128. doi: 10.1016/0022-2836(86)90298-6. [DOI] [PubMed] [Google Scholar]
  30. Stalder J., Reigel F., Koblet H. Defective viral RNAs in Aedes albopictus C6/36 cells persistently infected with Semliki Forest virus. Virology. 1983 Sep;129(2):247–254. doi: 10.1016/0042-6822(83)90164-2. [DOI] [PubMed] [Google Scholar]
  31. Wangh L. J., Osborne J. A., Hentschel C. C., Tilly R. Parenchymal cells purified from Xenopus liver and maintained in primary culture synthesize vitellogenin in response to estradiol-17 beta and serum albumin in response to dexamethasone. Dev Biol. 1979 Jun;70(2):479–499. doi: 10.1016/0012-1606(79)90040-x. [DOI] [PubMed] [Google Scholar]
  32. Widmer H. J., Andres A. C., Niessing J., Hosbach H. A., Weber R. Comparative analysis of cloned larval and adult globin cDNA sequences of Xenopus laevis. Dev Biol. 1981 Dec;88(2):325–332. doi: 10.1016/0012-1606(81)90176-7. [DOI] [PubMed] [Google Scholar]
  33. Widmer H. J., Hosbach H. A., Weber R. Globin gene expression in Xenopus laevis: anemia induces precocious globin transition and appearance of adult erythroblasts during metamorphosis. Dev Biol. 1983 Sep;99(1):50–60. doi: 10.1016/0012-1606(83)90253-1. [DOI] [PubMed] [Google Scholar]
  34. 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 Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES