Role of DNA sequences outside the cores of DNase hypersensitive sites (HSs) in functions of the β-globin locus control region. Domain opening and synergism between HS2 and HS3

John D. Jackson, Hanna Petrykowska, Sjaak Philipsen, Webb Miller, Ross Hardison

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

The roles of each DNase hypersensitive site (HS), and the DNA sequences between them, in the activity of the locus control region of the mammalian β-globin gene domain were examined by placing human and rabbit restriction fragments containing the cores of HS2, HS3, HS4, and HS5, along with varying amounts of flanking DNA, upstream of a hybrid ε-globin-luciferase reporter gene and testing for effects on expression both prior to and after integration into the chromosomes of K562 cells, a human erythroid cell line. Prior to integration, fragments containing HS2 enhanced expression to the greatest extent, and the modest enhancement by some fragments containing HS3 correlated with the presence of a well-conserved binding site for AP1/NFE2. The stronger effects of larger locus control region DNA fragments in clones of stably transfected cells indicates a role for sequences outside the HS cores after integration into the genome. The strong effect of a 1.9-kilobase HindIII fragment containing HS3 after, but not prior to, integration argues for the presence of a chromatin domain-opening activity. Use of a rabbit DNA fragment containing both HS2 and HS3 demonstrated a synergistic interaction between the two HSs when their natural context and spacing are preserved.

Original languageEnglish (US)
Pages (from-to)11871-11878
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number20
DOIs
StatePublished - 1996

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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