Home LiteratureArticle Details
PMID: 12941700 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

HS5 of the human beta-globin locus control region: a developmental stage-specific border in erythroid cells.

The EMBO journal ·Vol. 22 ·No. 17 ·2003-09-01 ·Pages 4489-500

Wai AW, Gillemans N, Raguz-Bolognesi S, Pruzina S, Zafarana G, Meijer D, Philipsen S, Grosveld F

Abstract

Elements with insulator/border activity have been characterized most extensively in Drosophila melanogaster. In vertebrates, the first example of such an element was provided by a hypersensitive site of the chicken beta-globin locus, cHS4. It has been proposed that the homologous site in humans, HS5, functions as a border of the human beta-globin locus. Here, we have characterized HS5 of the human beta-globin locus control region. We have examined its tissue-specificity and assessed its insulating properties in transgenic mice using a lacZ reporter assay. Most importantly, we have tested its enhancer blocking activity in the context of the full beta-globin locus. Our results show that HS5 is erythroid-specific rather than ubiquitous in human tissues. Furthermore, HS5 does not fulfil the criteria of a general in vivo insulator in the transgene protection assay. Finally, a HS5 conditional deletion from the complete locus demonstrates that HS5 has no discernable activity in adult erythroid cells. Surprisingly, HS5 functions as an enhancer blocker in embryonic erythroid cells. We conclude that HS5 is a developmental stage-specific border in erythroid cells.

MeSH Terms
Animals Chromosome Mapping Drosophila melanogaster/genetics Erythropoiesis/genetics Gene Expression Regulation, Developmental Globins/genetics Humans Lac Operon Locus Control Region Mice Mice, Transgenic Transcriptional Activation
Chemicals
Globins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Wai Albert W K
Department of Cell Biology, Erasmus MC, PO Box 1738, 3000 DR Rotterdam, The Netherlands.
Gillemans Nynke
Raguz-Bolognesi Selina
Pruzina Sara
Zafarana Gaetano
Meijer Dies
Philipsen Sjaak
Grosveld Frank
References (65)
65 references, click to expand
  1. Transitions in histone acetylation reveal boundaries of three separately regulated neighboring loci.
    EMBO J. 2001 May 1;20(9):2224-35 PMID: 11331588
  2. Intron-exon structure, alternative use of promoter and expression of the mouse collagen X gene, Col10a-1.
    Eur J Biochem. 1993 Apr 1;213(1):99-111 PMID: 8477738
  3. A position-effect assay for boundaries of higher order chromosomal domains.
    Cell. 1991 Mar 8;64(5):941-50 PMID: 1848159
  4. A deletion of the human beta-globin locus activation region causes a major alteration in chromatin structure and replication across the entire beta-globin locus.
    Genes Dev. 1990 Oct;4(10):1637-49 PMID: 2249769
  5. Heterochromatin effects on the frequency and duration of LCR-mediated gene transcription.
    Cell. 1996 Oct 4;87(1):105-14 PMID: 8858153
  6. The human beta-globin locus control region.
    Eur J Biochem. 2002 Mar;269(6):1589-99 PMID: 11895428
  7. Beta-globin gene switching and DNase I sensitivity of the endogenous beta-globin locus in mice do not require the locus control region.
    Mol Cell. 2000 Feb;5(2):387-93 PMID: 10882079
  8. Looping and interaction between hypersensitive sites in the active beta-globin locus.
    Mol Cell. 2002 Dec;10(6):1453-65 PMID: 12504019
  9. 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):2718-22 PMID: 6232614
  10. Position-independent, high-level expression of the human beta-globin gene in transgenic mice.
    Cell. 1987 Dec 24;51(6):975-85 PMID: 3690667
  11. The insulation of genes from external enhancers and silencing chromatin.
    Proc Natl Acad Sci U S A. 2002 Dec 10;99 Suppl 4:16433-7 PMID: 12154228
  12. Comparative kinetic analysis of FLP and cre recombinases: mathematical models for DNA binding and recombination.
    J Mol Biol. 1998 Nov 27;284(2):363-84 PMID: 9813124
  13. Long-range chromatin regulatory interactions in vivo.
    Nat Genet. 2002 Dec;32(4):623-6 PMID: 12426570
  14. The "beta-like-globin" gene domain in human erythroid cells.
    Proc Natl Acad Sci U S A. 1985 Oct;82(19):6384-8 PMID: 3879975
  15. Correlation between histone lysine methylation and developmental changes at the chicken beta-globin locus.
    Science. 2001 Sep 28;293(5539):2453-5 PMID: 11498546
  16. Erythroid Krüppel-like factor exhibits an early and sequentially localized pattern of expression during mammalian erythroid ontogeny.
    Dev Dyn. 1996 Jul;206(3):248-59 PMID: 8896981
  17. Functional gene expression domains: defining the functional unit of eukaryotic gene regulation.
    Bioessays. 2000 Jul;22(7):657-65 PMID: 10878578
  18. Erythroid Krüppel-like factor (EKLF) is active in primitive and definitive erythroid cells and is required for the function of 5'HS3 of the beta-globin locus control region.
    EMBO J. 1998 Apr 15;17(8):2334-41 PMID: 9545245
  19. An insulator element and condensed chromatin region separate the chicken beta-globin locus from an independently regulated erythroid-specific folate receptor gene.
    EMBO J. 1999 Jul 15;18(14):4035-48 PMID: 10406808
  20. Induction of Erythroid-Specific Expression in Murine Erythroleukemia (MEL) Cell Lines.
    Methods Mol Biol. 1991;7:421-34 PMID: 21416373
  21. Core histone hyperacetylation co-maps with generalized DNase I sensitivity in the chicken beta-globin chromosomal domain.
    EMBO J. 1994 Apr 15;13(8):1823-30 PMID: 8168481
  22. Transcription complex stability and chromatin dynamics in vivo.
    Nature. 1995 Sep 21;377(6546):209-13 PMID: 7675106
  23. Importance of globin gene order for correct developmental expression.
    Genes Dev. 1991 Aug;5(8):1387-94 PMID: 1714415
  24. A group of scs elements function as domain boundaries in an enhancer-blocking assay.
    Mol Cell Biol. 1992 May;12(5):2424-31 PMID: 1569958
  25. The role of EKLF in human beta-globin gene competition.
    Genes Dev. 1996 Nov 15;10(22):2894-902 PMID: 8918890
  26. The effect of distance on long-range chromatin interactions.
    Mol Cell. 1997 Dec;1(1):131-9 PMID: 9659910
  27. Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities.
    Proc Natl Acad Sci U S A. 2002 May 14;99(10):6883-8 PMID: 12011446
  28. Human CD2 3'-flanking sequences confer high-level, T cell-specific, position-independent gene expression in transgenic mice.
    Cell. 1989 Mar 24;56(6):979-86 PMID: 2564317
  29. Structural analysis and mapping of DNase I hypersensitivity of HS5 of the beta-globin locus control region.
    Genomics. 1999 Oct 15;61(2):183-93 PMID: 10534403
  30. Characterization of the chicken beta-globin insulator.
    Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):575-80 PMID: 9012826
  31. A transgenic mouse line that retains Cre recombinase activity in mature oocytes irrespective of the cre transgene transmission.
    Biochem Biophys Res Commun. 1997 Aug 18;237(2):318-24 PMID: 9268708
  32. Conserved CTCF insulator elements flank the mouse and human beta-globin loci.
    Mol Cell Biol. 2002 Jun;22(11):3820-31 PMID: 11997516
  33. The polyoma virus enhancer cannot substitute for DNase I core hypersensitive sites 2-4 in the human beta-globin LCR.
    Nucleic Acids Res. 1999 Aug 1;27(15):3130-7 PMID: 10454609
  34. The human beta-globin locus control region confers an early embryonic erythroid-specific expression pattern to a basic promoter driving the bacterial lacZ gene.
    Development. 1996 Dec;122(12):3991-9 PMID: 9012519
  35. 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):10159-77 PMID: 3480506
  36. A chromatin insulator protects retrovirus vectors from chromosomal position effects.
    Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):9150-5 PMID: 10908661
  37. Locus control regions of mammalian beta-globin gene clusters: combining phylogenetic analyses and experimental results to gain functional insights.
    Gene. 1997 Dec 31;205(1-2):73-94 PMID: 9461381
  38. Transcriptional regulation of vav, a gene expressed throughout the hematopoietic compartment.
    Blood. 1998 Jan 15;91(2):419-30 PMID: 9427694
  39. The beta-globin dominant control region: hypersensitive site 2.
    EMBO J. 1990 Jul;9(7):2159-67 PMID: 2357964
  40. Chromatin and gene regulation at the onset of embryonic development.
    Reprod Nutr Dev. 1996;36(6):581-606 PMID: 9021871
  41. Development of virus vectors for gene therapy of beta chain hemoglobinopathies: flanking with a chromatin insulator reduces gamma-globin gene silencing in vivo.
    Blood. 2002 Sep 15;100(6):2012-9 PMID: 12200360
  42. Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice.
    Development. 1989 Apr;105(4):707-14 PMID: 2557196
  43. The Fab-7 element of the bithorax complex attenuates enhancer-promoter interactions in the Drosophila embryo.
    Genes Dev. 1996 Dec 15;10(24):3195-201 PMID: 8985187
  44. Hypersensitive site 2 specifies a unique function within the human beta-globin locus control region to stimulate globin gene transcription.
    Mol Cell Biol. 1999 Apr;19(4):3062-72 PMID: 10082573
  45. Altered DNA-binding specificity mutants of EKLF and Sp1 show that EKLF is an activator of the beta-globin locus control region in vivo.
    Genes Dev. 1998 Sep 15;12(18):2863-73 PMID: 9744863
  46. Hypersensitive site 5 of the human beta locus control region functions as a chromatin insulator.
    Blood. 1994 Sep 1;84(5):1399-401 PMID: 8068937
  47. A long terminal repeat of the human endogenous retrovirus ERV-9 is located in the 5' boundary area of the human beta-globin locus control region.
    Genomics. 1998 Dec 15;54(3):542-55 PMID: 9878258
  48. An in vitro globin gene switching model based on differentiated embryonic stem cells.
    Genes Dev. 1990 Dec;4(12A):2075-85 PMID: 2269427
  49. Serial deletions and duplications suggest a mechanism for the collinearity of Hoxd genes in limbs.
    Nature. 2002 Nov 14;420(6912):145-50 PMID: 12432383
  50. Each hypersensitive site of the human beta-globin locus control region confers a different developmental pattern of expression on the globin genes.
    Genes Dev. 1993 Jan;7(1):106-13 PMID: 8422981
  51. The beta-globin LCR is not necessary for an open chromatin structure or developmentally regulated transcription of the native mouse beta-globin locus.
    Mol Cell. 1998 Oct;2(4):447-55 PMID: 9809066
  52. The su(Hw) protein insulates expression of the Drosophila melanogaster white gene from chromosomal position-effects.
    EMBO J. 1993 Feb;12(2):435-42 PMID: 8382607
  53. Intergenic transcription and developmental remodeling of chromatin subdomains in the human beta-globin locus.
    Mol Cell. 2000 Feb;5(2):377-86 PMID: 10882078
  54. Evidence that DNase I hypersensitive site 5 of the human beta-globin locus control region functions as a chromosomal insulator in transgenic mice.
    Nucleic Acids Res. 2002 Jun 1;30(11):2484-91 PMID: 12034837
  55. LCR/MEL: a versatile system for high-level expression of heterologous proteins in erythroid cells.
    Nucleic Acids Res. 1992 Mar 11;20(5):997-1003 PMID: 1549512
  56. The locus control region is necessary for gene expression in the human beta-globin locus but not the maintenance of an open chromatin structure in erythroid cells.
    Mol Cell Biol. 1998 Oct;18(10):5992-6000 PMID: 9742116
  57. Developmental regulation of a complete 70-kb human beta-globin locus in transgenic mice.
    Genes Dev. 1992 Oct;6(10):1857-64 PMID: 1383089
  58. Beta-globin gene inactivation by DNA translocation in gamma beta-thalassaemia.
    Nature. 1983 Dec 15-21;306(5944):662-6 PMID: 6318113
  59. Detailed analysis of the site 3 region of the human beta-globin dominant control region.
    EMBO J. 1990 Jul;9(7):2169-77 PMID: 2357965
  60. A 5' element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects against position effect in Drosophila.
    Cell. 1993 Aug 13;74(3):505-14 PMID: 8348617
  61. Role of gene order in developmental control of human gamma- and beta-globin gene expression.
    Mol Cell Biol. 1993 Aug;13(8):4836-43 PMID: 8336720
  62. Effects of altered gene order or orientation of the locus control region on human beta-globin gene expression in mice.
    Nature. 1999 Mar 25;398(6725):344-8 PMID: 10192336
  63. Loss of transcriptional activity of a transgene is accompanied by DNA methylation and histone deacetylation and is prevented by insulators.
    Genes Dev. 1998 Sep 15;12(18):2852-62 PMID: 9744862
  64. The minimal requirements for activity in transgenic mice of hypersensitive site 3 of the beta globin locus control region.
    EMBO J. 1993 Mar;12(3):1077-85 PMID: 8458325
  65. Modification of human beta-globin locus PAC clones by homologous recombination in Escherichia coli.
    Nucleic Acids Res. 2000 Jun 15;28(12):E65 PMID: 10871388
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg437
Published
2003-09-01
Pages
4489-500
Language
English
Region
England
NLM ID
8208664
PMCID
PMC202379
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/cdg437 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2