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An Erythroid-Specific Chromatin Opening Element Reorganizes β-Globin Promoter Chromatin Structure and Augments Gene Expression

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Abstract

ABSTRACT

In erythroid tissues the chromatin structure of the β-globin gene locus is extensively remodeled. Changes include the formation of DNase I hypersensitive sites (HSs) over the promoters of actively expressed genes. To test the hypothesis that such “opening” of promoter chromatin structure is important for β-globin gene expression, we placed a 101-bp erythroid-specific hypersensitive-site forming element (HSFE) from the core of LCR HS4 immediately upstream of a minimal β-globin gene promoter. We then studied the effects of this element alone and in combination with other cis-acting elements on globin gene chromatin structure and gene expression in MEL cells and transgenic mice. Single or tandem HSFEs increased the size of the portion of the promoter accessible to DNase digestion, increased the proportion of promoters in an accessible conformation, and increased gene expression approximately 5-fold. These were equivalent to expression levels attained using a 2.8-kb μLCR construct. Inclusion of the LCR HS2 enhancer did not increase expression further. In transgenic mouse fetal liver cells the HSFE increased average expression 2.5-fold compared to the minimal promoter alone. These results indicate that a small cis-acting element is capable of remodeling local β-globin promoter chromatin structure and producing expression similar to that seen with a μLCR construct.

References (48)

  • D. Kioussis et al.

    Beta-globin gene inactivation by DNA translocation in γβ-thalassemia

    Nature

    (1983)
  • W.C. Forrester et al.

    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)
  • M. Groudine et al.

    Human fetal to adult hemoglobin switching: changes in chromatin structure of the β-globin gene locus

    Proc. Natl. Acad. Sci. USA

    (1983)
  • D. Tuan et al.

    The “beta-like-globin” gene domain in human erythroid cells

    Proc. Natl. Acad. Sci. USA

    (1985)
  • V. Dhar et al.

    Erythroid-specific nuclease-hypersensitive sites flanking the human β-globin domain

    Mol. Cell. Biol.

    (1990)
  • W.C. Forrester et al.

    Evidence for a locus activation region: The formation of developmentally-stable hypersensitive sites in globin expressing hybrids

    Nucleic Acids Res.

    (1987)
  • A. Reik et al.

    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)
  • M. Driscoll et al.

    γδβ thalassemia due to a de novo mutation deleting the 5′β-globin gene activation region hypersensitive sites

    Proc. Natl. Acad. Sci. USA

    (1989)
  • P. Curtin et al.

    A distant gene deletion affects β-globin in an atypical γδβ thalassemia

    J. Clin. Invest.

    (1985)
  • G. Felsenfeld

    Chromatin as an essential part of the transcriptional mechanism

    Nature

    (1992)
  • J. Bungert et al.

    Synergistic regulation of human beta-globin gene switching by locus control region elements HS3 and HS4

    Genes Dev.

    (1995)
  • D. Clark et al.

    Chromatin structure of transcriptionally active genes

    Cold Spring Harb. Symp. Quant. Biol.

    (1993)
  • Q. Gong et al.

    Essential role of NF-E2 in remodeling of chromatin structure and transcriptional activation of the epsilon-globin gene in vivo by 5′ hypersensitive site 2 of the beta-globin locus control region

    Mol. Cell. Biol.

    (1996)
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      DNase I Hypersensitive Site Mapping—DNase I hypersensitive site mapping was performed as described with minor modifications (27, 28).

    Communicated by G. Stamatoyannopoulos, M.D., 08/06/01

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    Correspondence and reprint requests to: Christopher H. Lowrey, Department of Medicine, Hinman Box 7650, Dartmouth Medical School, Hanover, NH 03755. Fax: (603) 650-1129. E-mail: [email protected].

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