Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1990 Mar;10(3):1209–1216. doi: 10.1128/mcb.10.3.1209

Negative regulation of the human epsilon-globin gene by transcriptional interference: role of an Alu repetitive element.

J Wu 1, G J Grindlay 1, P Bushel 1, L Mendelsohn 1, M Allan 1
PMCID: PMC360999  PMID: 2304465

Abstract

The human epsilon-globin gene has a number of alternative transcription initiation sites which correspond with regions of DNase I hypersensitivity upstream of the canonical cap site. Transcripts originating from the promoters located -4.3/-4.5 and -1.48 kilobase pairs (kbp) and -900 and -200 base pairs (bp) upstream of the major epsilon-globin cap site can, at certain stages of erythroid differentiation, extend through the gene and are polyadenylated. The 350-bp PolIII transcripts, originating within the Alu repetitive element -2.2 kbp upstream of the cap site, extend in the opposite direction from the gene, are nonpolyadenylated, nucleus confined, and are detectable only in mature K562 cells or mature embryonic red blood cells where the epsilon-globin major cap site is maximally transcribed. Fragments containing the promoters located between -4.5 and -4.3 kbp upstream of the gene down regulate transcription from the epsilon-globin gene 20- to 30-fold in a transient expression assay in which both erythroid and nonerythroid cell lines were used. This occurs only when the direction of transcription from the -4.3/-4.5-kbp promoters is towards the gene, and we hypothesize that down regulation is caused by transcriptional interference. Fragments containing the Alu repetitive element -2.2 kbp upstream of the gene can overcome down regulation of the epsilon-globin gene by the -4.5-kbp element when interposed in the direct orientation between this element and the epsilon-globin gene.

Full text

PDF
1216

Images in this article

1212Image
on p.1212
1213Image
on p.1213
1213Image
on p.1213
1214Image
on p.1214

Selected References

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

  1. Alan M., Grindlay G. J., Stefani L., Paul J. Epsilon globin gene transcripts originating upstream of the mRNA cap site in K562 cells and normal human embryos. Nucleic Acids Res. 1982 Sep 11;10(17):5133–5147. doi: 10.1093/nar/10.17.5133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allan M., Lanyon W. G., Paul J. Multiple origins of transcription in the 4.5 Kb upstream of the epsilon-globin gene. Cell. 1983 Nov;35(1):187–197. doi: 10.1016/0092-8674(83)90221-0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Allan M., Paul J. Transcription in vivo of an Alu family member upstream from the human epsilon-globin gene. Nucleic Acids Res. 1984 Jan 25;12(2):1193–1200. doi: 10.1093/nar/12.2.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Allan M., Zhu J. D., Montague P., Paul J. Differential response of multiple epsilon-globin cap sites to cis- and trans-acting controls. Cell. 1984 Sep;38(2):399–407. doi: 10.1016/0092-8674(84)90495-1. [DOI] [PubMed] [Google Scholar]
  6. Bateman E., Paule M. R. Promoter occlusion during ribosomal RNA transcription. Cell. 1988 Sep 23;54(7):985–992. doi: 10.1016/0092-8674(88)90113-4. [DOI] [PubMed] [Google Scholar]
  7. Bushel P., Rego K., Mendelsohn L., Allan M. Correlation between patterns of DNase I-hypersensitive sites and upstream promoter activity of the human epsilon-globin gene at different stages of erythroid development. Mol Cell Biol. 1990 Mar;10(3):1199–1208. doi: 10.1128/mcb.10.3.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cao S. X., Gutman P. D., Dave H. P., Schechter A. N. Identification of a transcriptional silencer in the 5'-flanking region of the human epsilon-globin gene. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5306–5309. doi: 10.1073/pnas.86.14.5306. [DOI] [PMC free article] [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. Coggins L. W., Lanyon W. G., Slater A. A., Grindlay G. J., Paul J. Characterization of Alu family repetitive sequences which flank human beta-type globin genes. Biosci Rep. 1981 Apr;1(4):309–317. doi: 10.1007/BF01114870. [DOI] [PubMed] [Google Scholar]
  11. Contreras R., Gheysen D., Knowland J., van de Voorde A., Fiers W. Evidence for the direct involvement of DNA replication origin in synthesis of late SV40 RNA. Nature. 1982 Dec 9;300(5892):500–505. doi: 10.1038/300500a0. [DOI] [PubMed] [Google Scholar]
  12. Corbin V., Maniatis T. Role of transcriptional interference in the Drosophila melanogaster Adh promoter switch. Nature. 1989 Jan 19;337(6204):279–282. doi: 10.1038/337279a0. [DOI] [PubMed] [Google Scholar]
  13. Cullen B. R., Lomedico P. T., Ju G. Transcriptional interference in avian retroviruses--implications for the promoter insertion model of leukaemogenesis. Nature. 1984 Jan 19;307(5948):241–245. doi: 10.1038/307241a0. [DOI] [PubMed] [Google Scholar]
  14. Di Segni G., Carrara G., Tocchini-Valentini G. R., Shoulders C. C., Baralle F. E. Selective in vitro transcription of one of the two Alu family repeats present in the 5' flanking region of the human epsilon-globin gene. Nucleic Acids Res. 1981 Dec 21;9(24):6709–6722. doi: 10.1093/nar/9.24.6709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dynan W. S., Tjian R. Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II. Cell. 1983 Mar;32(3):669–680. doi: 10.1016/0092-8674(83)90053-3. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Fox G. M., Hess J. F., Shen C. K., Schmid C. W. Alu family members in the human alpha-like globin-gene cluster. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):1131–1134. doi: 10.1101/sqb.1983.047.01.127. [DOI] [PubMed] [Google Scholar]
  18. Grindlay G. J., Lanyon W. G., Allan M., Paul J. Alternative sites of transcription initiation upstream of the canonical cap site in human gamma-globin and beta-globin genes. Nucleic Acids Res. 1984 Feb 24;12(4):1811–1820. doi: 10.1093/nar/12.4.1811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grosveld F., van Assendelft G. B., Greaves D. R., Kollias G. Position-independent, high-level expression of the human beta-globin gene in transgenic mice. Cell. 1987 Dec 24;51(6):975–985. doi: 10.1016/0092-8674(87)90584-8. [DOI] [PubMed] [Google Scholar]
  20. Harrison P. R. Analysis of erythropoeisis at the molecular level. Nature. 1976 Jul 29;262(5567):353–356. doi: 10.1038/262353a0. [DOI] [PubMed] [Google Scholar]
  21. Jelinek W. R., Schmid C. W. Repetitive sequences in eukaryotic DNA and their expression. Annu Rev Biochem. 1982;51:813–844. doi: 10.1146/annurev.bi.51.070182.004121. [DOI] [PubMed] [Google Scholar]
  22. Jones K. A., Kadonaga J. T., Rosenfeld P. J., Kelly T. J., Tjian R. A cellular DNA-binding protein that activates eukaryotic transcription and DNA replication. Cell. 1987 Jan 16;48(1):79–89. doi: 10.1016/0092-8674(87)90358-8. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Moss T. A transcriptional function for the repetitive ribosomal spacer in Xenopus laevis. Nature. 1983 Mar 17;302(5905):223–228. doi: 10.1038/302223a0. [DOI] [PubMed] [Google Scholar]
  25. Nelson K. J., Haimovich J., Perry R. P. Characterization of productive and sterile transcripts from the immunoglobulin heavy-chain locus: processing of micron and muS mRNA. Mol Cell Biol. 1983 Jul;3(7):1317–1332. doi: 10.1128/mcb.3.7.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Osborne T. F., Berk A. J. Far upstream initiation sites for adenovirus early region 1A transcription are utilized after the onset of viral DNA replication. J Virol. 1983 Feb;45(2):594–599. doi: 10.1128/jvi.45.2.594-599.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Page G. S., Smith S., Goodman H. M. DNA sequence of the rat growth hormone gene: location of the 5' terminus of the growth hormone mRNA and identification of an internal transposon-like element. Nucleic Acids Res. 1981 May 11;9(9):2087–2104. doi: 10.1093/nar/9.9.2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Proudfoot N. J. Transcriptional interference and termination between duplicated alpha-globin gene constructs suggests a novel mechanism for gene regulation. Nature. 1986 Aug 7;322(6079):562–565. doi: 10.1038/322562a0. [DOI] [PubMed] [Google Scholar]
  29. Rohrbaugh M. L., Johnson J. E., 3rd, James M. D., Hardison R. C. Transcription unit of the rabbit beta 1 globin gene. Mol Cell Biol. 1985 Jan;5(1):147–160. doi: 10.1128/mcb.5.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Tuan D., London I. M. Mapping of DNase I-hypersensitive sites in the upstream DNA of human embryonic epsilon-globin gene in K562 leukemia cells. Proc Natl Acad Sci U S A. 1984 May;81(9):2718–2722. doi: 10.1073/pnas.81.9.2718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  33. Wu C. Two protein-binding sites in chromatin implicated in the activation of heat-shock genes. Nature. 1984 May 17;309(5965):229–234. doi: 10.1038/309229a0. [DOI] [PubMed] [Google Scholar]
  34. Young P. R., Scott R. W., Hamer D. H., Tilghman S. M. Construction and expression in vivo of an internally deleted mouse alpha-fetoprotein gene: presence of a transcribed Alu-like repeat within the first intervening sequence. Nucleic Acids Res. 1982 May 25;10(10):3099–3116. doi: 10.1093/nar/10.10.3099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zhu J., Allan M., Paul J. The chromatin structure of the human epsilon globin gene: nuclease hypersensitive sites correlate with multiple initiation sites of transcription. Nucleic Acids Res. 1984 Dec 11;12(23):9191–9204. doi: 10.1093/nar/12.23.9191. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES