FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter

Susan K. Durham, Adisak Suwanichkul, Ann Scheimann, Douglas Yee, James G. Jackson, Frederic G. Barr, David R. Powell

Research output: Contribution to journalArticle

Abstract

The insulin response element (IRE) in the IGFBP-1 promoter, and in other gene promoters, contains a T(A/G)TTT motif essential for insulin inhibition of transcription. Studies presented here test whether FKHR may be the transcription factor that confers insulin inhibition through this IRE motif. Immunoblots using antiserum to the synthetic peptide FKHR413-480, RNase protection, and Northerns blots show that FKHR is expressed in HEP G2 human hepatoma cells. Southwestern blots, electromobility shift assays, and DNase I protection assays show that Escherichia coli-expressed GST-FKHR binds specifically to IREs from the IGFBP-1, PEPCK and TAT genes; however, unlike HNF3β, another protein proposed to be the insulin regulated factor, GST-FKHR does not bind the insulin unresponsive G/C-A/C mutation of the IGFBP-1 IRE. When HEP G2 cells were cotransfected with FKHR expression vectors and with IGFBP-1 promoter plasmids containing either native or mutant IREs, FKHR expression induced a 5-fold increase in activity of the native IGFBP-1 promoter but no increase in activity of promoter constructs containing insulin unresponsive IRE mutants. These data suggest that FKHR, and/or a related family member, is the important T(G/A)TTT binding protein that confers the inhibitory effect of insulin on gene transcription.

Original languageEnglish (US)
Pages (from-to)3140-3146
Number of pages7
JournalEndocrinology
Volume140
Issue number7
StatePublished - 1999
Externally publishedYes

Fingerprint

Insulin-Like Growth Factor Binding Protein 1
Response Elements
Insulin
Southwestern Blotting
Genes
Deoxyribonuclease I
Ribonucleases
Immune Sera
Hepatocellular Carcinoma
Carrier Proteins
Plasmids
Transcription Factors
Escherichia coli

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Durham, S. K., Suwanichkul, A., Scheimann, A., Yee, D., Jackson, J. G., Barr, F. G., & Powell, D. R. (1999). FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter. Endocrinology, 140(7), 3140-3146.

FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter. / Durham, Susan K.; Suwanichkul, Adisak; Scheimann, Ann; Yee, Douglas; Jackson, James G.; Barr, Frederic G.; Powell, David R.

In: Endocrinology, Vol. 140, No. 7, 1999, p. 3140-3146.

Research output: Contribution to journalArticle

Durham, SK, Suwanichkul, A, Scheimann, A, Yee, D, Jackson, JG, Barr, FG & Powell, DR 1999, 'FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter', Endocrinology, vol. 140, no. 7, pp. 3140-3146.
Durham SK, Suwanichkul A, Scheimann A, Yee D, Jackson JG, Barr FG et al. FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter. Endocrinology. 1999;140(7):3140-3146.
Durham, Susan K. ; Suwanichkul, Adisak ; Scheimann, Ann ; Yee, Douglas ; Jackson, James G. ; Barr, Frederic G. ; Powell, David R. / FKHR binds the insulin response element in the insulin-like growth factor binding protein-1 promoter. In: Endocrinology. 1999 ; Vol. 140, No. 7. pp. 3140-3146.
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