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Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting.

Publication ,  Journal Article
Cameron, S; Taylor, DS; TePas, EC; Speck, NA; Mathey-Prevot, B
Published in: Blood
May 15, 1994

Interleukin-3 (IL-3) is involved in proliferation and differentiation of hematopoietic progenitor cells. Its expression is subject to precise, tissue-specific regulation, and has been studied extensively in the gibbon T-cell line MLA 144 by a combination of functional assays and DNA binding experiments. To extend these studies, the gibbon IL-3 promoter was cloned and in vivo footprinting of the gibbon and human IL-3 proximal promoters was performed. The gibbon IL-3 promoter was found to be highly homologous to its human counterpart and both promoters yielded identical in vivo footprints after induction of IL-3 synthesis. In particular, we observed specific protection of three guanines over a core sequence TGTGGTTT (IF-1IL3) that had not been recognized in previous studies. IF-1IL3 is not found in other cytokine promoters, but it is conserved in the IL-3 promoter of several species and is similar to a recurring motif in viral and T-cell-specific cellular enhancers. IF-1IL3 binds a specific complex in MLA 144 and Jurkat nuclear extracts in vitro, which shares the same specificity as the complex bound by the polyoma virus and T-cell receptor delta enhancers. Mutation of the three guanines in IF-1IL3 core sequence disrupts binding in vitro and abrogates the ability of the IL-3 promoter to mediate inducible expression in T cells. Although IF-1IL3 is necessary for IL-3 expression, it is not sufficient: a truncated IL-3 promoter with an intact IF-1IL3 site but no other activator sites is transcriptionally silent. These studies describe a new regulatory element within the IL-3 promoter that is essential for expression and conserved between species.

Duke Scholars

Published In

Blood

ISSN

0006-4971

Publication Date

May 15, 1994

Volume

83

Issue

10

Start / End Page

2851 / 2859

Location

United States

Related Subject Headings

  • Receptors, Antigen, T-Cell, gamma-delta
  • Promoter Regions, Genetic
  • Molecular Sequence Data
  • Interleukin-3
  • Immunology
  • Humans
  • Genes, Regulator
  • Cell Line
  • Base Sequence
  • Amino Acid Sequence
 

Citation

APA
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MLA
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Cameron, S., Taylor, D. S., TePas, E. C., Speck, N. A., & Mathey-Prevot, B. (1994). Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting. Blood, 83(10), 2851–2859.
Cameron, S., D. S. Taylor, E. C. TePas, N. A. Speck, and B. Mathey-Prevot. “Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting.Blood 83, no. 10 (May 15, 1994): 2851–59.
Cameron S, Taylor DS, TePas EC, Speck NA, Mathey-Prevot B. Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting. Blood. 1994 May 15;83(10):2851–9.
Cameron, S., et al. “Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting.Blood, vol. 83, no. 10, May 1994, pp. 2851–59.
Cameron S, Taylor DS, TePas EC, Speck NA, Mathey-Prevot B. Identification of a critical regulatory site in the human interleukin-3 promoter by in vivo footprinting. Blood. 1994 May 15;83(10):2851–2859.

Published In

Blood

ISSN

0006-4971

Publication Date

May 15, 1994

Volume

83

Issue

10

Start / End Page

2851 / 2859

Location

United States

Related Subject Headings

  • Receptors, Antigen, T-Cell, gamma-delta
  • Promoter Regions, Genetic
  • Molecular Sequence Data
  • Interleukin-3
  • Immunology
  • Humans
  • Genes, Regulator
  • Cell Line
  • Base Sequence
  • Amino Acid Sequence