In vivo protein-DNA interactions at the β-globin gene locus

Tohru Ikuta, Yuet Wai Kan

Research output: Contribution to journalArticlepeer-review

91 Scopus citations

Abstract

We have investigated in vivo protein-DNA interactions in the β-globin gene locus by dimethyl sulfate (DMS) footprinting in K562 cells, which express ε- and γ-globin but not β-globin. In the locus control region, hypersensitive site 2 (HS-2) exhibited footprints in several putative protein binding motifs. HS-3 was not footprinted. The β promoter was also not footprinted, while extensive footprints were observed in the promoter of the active γ-globin gene. No footprints were seen in the Aγ and β3′ enhancers. With several motifs, additional protein interactions and alterations in binding patterns occurred with hemin induction. In HeLa cells, some footprints were observed in some of the motifs in HS-2, compatible with the finding that HS-2 has some enhancer function in HeLa cells, albeit much weaker than its activity in K562 cells. No footprint was seen in B lymphocytes. In vivo footprinting is a useful method for studying relevant protein-DNA interactions in erythroid cells.

Original languageEnglish (US)
Pages (from-to)10188-10192
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number22
StatePublished - 1991
Externally publishedYes

Keywords

  • B lymphocyte
  • Dimethyl sulfate
  • HeLa cells
  • In vivo footprinting
  • K562 cells

ASJC Scopus subject areas

  • General

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