Alterations in protein-DNA interactions in the γ-globin gene promoter in response to butyrate therapy

Tohru Ikuta, Yuet Wai Kan, Paul S. Swerdlow, Douglas V. Faller, Susan P. Perrine

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

The mechanisms by which pharmacologic agents stimulate γ-globin gene expression in β-globin disorders has not been fully established at the molecular level. In studies described here, nucleated erythroblasts were isolated from patients with β-globin disorders before and with butyrate therapy, and globin biosynthesis, mRNA, and protein-DNA interactions were examined. Expression of γ-globin mRNA increased twofold to sixfold above baseline with butyrate therapy in 7 of 8 patients studied. A 15% to 50% increase in γ-globin protein synthetic levels above baseline γ globin ratios and a relative decrease in β-globin biosynthesis were observed in responsive patients. Extensive new in vivo footprints were detected in erythroblasts of responsive patients in four regions of the γ-globin gene promoter, designated butyrate-response elements gamma 1-4 (BRE-G1-4). Electrophoretic mobility shift assays using BRE-G1 sequences as a probe demonstrated that new binding of two erythroid-specific proteins and one ubiquitous protein, αCP2, occurred with treatment in the responsive patients and did not occur in the nonresponder. The BRE-G1 sequence conferred butyrate inducibility in reporter gene assays. These in vivo protein-DNA interactions in human erythroblasts in which γ-globin gene expression is being altered strongly suggest that nuclear protein binding, including αCP2, to the BRE- G1 region of the γ-globin gene promoter mediates butyrate activity on γ- globin gene expression.

Original languageEnglish (US)
Pages (from-to)2924-2933
Number of pages10
JournalBlood
Volume92
Issue number8
StatePublished - Oct 15 1998
Externally publishedYes

Fingerprint

Globins
Butyrates
Genes
DNA
Proteins
Erythroblasts
Therapeutics
Gene expression
Biosynthesis
Gene Expression
Assays
Electrophoretic mobility
Messenger RNA
Protein Biosynthesis
Response Elements
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Reporter Genes
Protein Binding

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Ikuta, T., Kan, Y. W., Swerdlow, P. S., Faller, D. V., & Perrine, S. P. (1998). Alterations in protein-DNA interactions in the γ-globin gene promoter in response to butyrate therapy. Blood, 92(8), 2924-2933.

Alterations in protein-DNA interactions in the γ-globin gene promoter in response to butyrate therapy. / Ikuta, Tohru; Kan, Yuet Wai; Swerdlow, Paul S.; Faller, Douglas V.; Perrine, Susan P.

In: Blood, Vol. 92, No. 8, 15.10.1998, p. 2924-2933.

Research output: Contribution to journalArticle

Ikuta, T, Kan, YW, Swerdlow, PS, Faller, DV & Perrine, SP 1998, 'Alterations in protein-DNA interactions in the γ-globin gene promoter in response to butyrate therapy', Blood, vol. 92, no. 8, pp. 2924-2933.
Ikuta T, Kan YW, Swerdlow PS, Faller DV, Perrine SP. Alterations in protein-DNA interactions in the γ-globin gene promoter in response to butyrate therapy. Blood. 1998 Oct 15;92(8):2924-2933.
Ikuta, Tohru ; Kan, Yuet Wai ; Swerdlow, Paul S. ; Faller, Douglas V. ; Perrine, Susan P. / Alterations in protein-DNA interactions in the γ-globin gene promoter in response to butyrate therapy. In: Blood. 1998 ; Vol. 92, No. 8. pp. 2924-2933.
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