Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 Sep 25;14(18):7199–7212. doi: 10.1093/nar/14.18.7199

Beta-globin gene promoter generates 5' truncated transcripts in the embryonic/fetal erythroid environment.

K Khazaie, F Gounari, M Antoniou, E deBoer, F Grosveld
PMCID: PMC311746  PMID: 2429259

Abstract

We report here the localisation of sequences responsible for the faulty expression of human beta-globin gene in Putko and K562 cells. Complete beta-globin gene introduced into these cells produces transcripts with abnormal 5' ends, while cotransfected mouse H2 gene is expressed correctly. Using hybrid constructs of these two genes we demonstrate that aberrant activity is conferred by sequences 5' of the beta-globin gene. Thus beta-globin promoter attached to the H2 coding sequence produces H2 transcripts with truncated 5' ends. By introducing a series of deletions in the beta-globin promoter we restrict these sequences to the -77/+28 base pair region spanning the CAAT element to the translation initiation site. These results are consistent with the lack of recognition of the beta-globin gene major cap site in Putko and K562 cells. We suggest that inactivity of the adult globin gene in the embryonic/fetal environment is at least in part conferred by sequences within the beta-globin gene promoter.

Full text

PDF
7199

Images in this article

7202Image
on p.7202
7203Image
on p.7203
7204Image
on p.7204
7206Image
on p.7206
7208Image
on p.7208
7209Image
on p.7209

Selected References

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

  1. Allan M., Montague P., Grindlay G. J., Sibbet G., Donovan-Peluso M., Bank A., Paul J. Tissue specific transcription of the human epsilon-globin gene following transfection into the embryonic erythroid cell line K562. Nucleic Acids Res. 1985 Sep 11;13(17):6125–6136. doi: 10.1093/nar/13.17.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anagnou N. P., Karlsson S., Moulton A. D., Keller G., Nienhuis A. W. Promoter sequences required for function of the human gamma globin gene in erythroid cells. EMBO J. 1986 Jan;5(1):121–126. doi: 10.1002/j.1460-2075.1986.tb04185.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chada K., Magram J., Raphael K., Radice G., Lacy E., Costantini F. Specific expression of a foreign beta-globin gene in erythroid cells of transgenic mice. 1985 Mar 28-Apr 3Nature. 314(6009):377–380. doi: 10.1038/314377a0. [DOI] [PubMed] [Google Scholar]
  4. Charnay P., Maniatis T. Transcriptional regulation of globin gene expression in the human erythroid cell line K562. Science. 1983 Jun 17;220(4603):1281–1283. doi: 10.1126/science.6574602. [DOI] [PubMed] [Google Scholar]
  5. Charnay P., Mellon P., Maniatis T. Linker scanning mutagenesis of the 5'-flanking region of the mouse beta-major-globin gene: sequence requirements for transcription in erythroid and nonerythroid cells. Mol Cell Biol. 1985 Jun;5(6):1498–1511. doi: 10.1128/mcb.5.6.1498. [DOI] [PMC free article] [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. 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]
  8. 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]
  9. Grosveld F. G., Lund T., Murray E. J., Mellor A. L., Dahl H. H., Flavell R. A. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 1982 Nov 11;10(21):6715–6732. doi: 10.1093/nar/10.21.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harrison P. R., Conkie D., Affara N., Paul J. In situ localization of globin messenger RNA formation. I. During mouse fetal liver development. J Cell Biol. 1974 Nov;63(2 Pt 1):402–413. doi: 10.1083/jcb.63.2.402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kioussis D., Wilson F., Khazaie K., Grosveld F. Differential expression of human globin genes introduced in K562 cells. EMBO J. 1985 Apr;4(4):927–931. doi: 10.1002/j.1460-2075.1985.tb03720.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klein G., Zeuthen J., Eriksson I., Terasaki P., Bernoco M., Rosén A., Masucci G., Povey S., Ber R. Hybridization of a myeloid leukemia-derived human cell line (K562) with a human Burkitt's lymphoma line (P3HR-1). J Natl Cancer Inst. 1980 Apr;64(4):725–738. [PubMed] [Google Scholar]
  13. 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]
  14. Magram J., Chada K., Costantini F. Developmental regulation of a cloned adult beta-globin gene in transgenic mice. Nature. 1985 May 23;315(6017):338–340. doi: 10.1038/315338a0. [DOI] [PubMed] [Google Scholar]
  15. Maniatis T., Fritsch E. F., Lauer J., Lawn R. M. The molecular genetics of human hemoglobins. Annu Rev Genet. 1980;14:145–178. doi: 10.1146/annurev.ge.14.120180.001045. [DOI] [PubMed] [Google Scholar]
  16. Marks P. A., Rifkind R. A. Erythroleukemic differentiation. Annu Rev Biochem. 1978;47:419–448. doi: 10.1146/annurev.bi.47.070178.002223. [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. 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]
  19. Townes T. M., Lingrel J. B., Chen H. Y., Brinster R. L., Palmiter R. D. Erythroid-specific expression of human beta-globin genes in transgenic mice. EMBO J. 1985 Jul;4(7):1715–1723. doi: 10.1002/j.1460-2075.1985.tb03841.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]
  23. Young K., Donovan-Peluso M., Cubbon R., Bank A. Trans acting regulation of beta globin gene expression in erythroleukemia (K562) cells. Nucleic Acids Res. 1985 Jul 25;13(14):5203–5213. doi: 10.1093/nar/13.14.5203. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES