Insulin inhibits transcription of the human gene for insulin-like growth factor-binding protein-1

David R. Powell, Adisak Suwanichkul, Michael L. Cubbage, Laura A. DePaolis, Mark B. Snuggs, Phillip Lee

Research output: Contribution to journalArticle

180 Citations (Scopus)

Abstract

Insulin rapidly lowers serum insulin-like growth factor-binding protein-1 (IGFBP-1) levels in vivo. In studies reported here, HEP G2 cells were used as a model system to investigate how insulin achieves this effect. When HEP G2 cells were incubated with 100 nM insulin for 6, 14, or 24 h, IGFBP-1 protein levels in conditioned medium fell to ∼50% of control values. This apparently was due to a fall in the rate of IGFBP-1 protein synthesis, since HEP G2 cells incorporated 46% less [35S]methionine into IGFBP-1 during a 4-h incubation with 100 nM insulin. IGFBP-1 mRNA levels were similarly affected by 100 nM insulin, falling to 45% of control values after 2 h, and to 9% of control values after 4 h of incubation with this hormone. The fall in IGFBP-1 mRNA level is consistent with data from nuclear transcription assays. HEP G2 nuclei isolated from cells that were incubated with 100 nM insulin for 2 h synthesized only ∼1/3 the number of IGFBP-1 transcripts as did control nuclei. Further evidence that insulin decreases IGFBP-1 gene transcription comes from transient transfections using chimeric IGFBP-1 promoter-chloramphenicol acetyltransferase reporter gene constructs. IGFBP-1 promoter activity fell to ∼50% of control values when HEP G2 cells transfected with a construct containing the first 1205 base pairs of the IGFBP-1 promoter were incubated with 100 nM insulin for 6, 14, or 24 h. Insulin lowered both IGFBP-1 protein levels and promoter activity in a dose-dependent manner. A half-maximal effect was found at ∼1 nM insulin and a maximal effect was found at ∼10 nM insulin in each instance. Transfections with constructs containing smaller IGFBP-1 promoter fragments showed that the region spanning from 103 to 529 base pairs 5′ to the IGFBP-1 mRNA cap site was necessary to demonstrate the inhibitory effect of insulin. These studies indicate that insulin lowers IGFBP-1 protein levels, at least in part, by rapidly decreasing the rate of IGFBP-1 gene transcription, and suggest that this insulin-mediated fall in transcription is conferred through a specific region of the IGFBP-1 promoter.

Original languageEnglish (US)
Pages (from-to)18868-18876
Number of pages9
JournalJournal of Biological Chemistry
Volume266
Issue number28
StatePublished - Oct 5 1991
Externally publishedYes

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Insulin-Like Growth Factor Binding Protein 1
Transcription
Genes
Insulin
human IGFBP1 protein
Base Pairing
Messenger RNA
Transfection
Proteins
Chloramphenicol O-Acetyltransferase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Powell, D. R., Suwanichkul, A., Cubbage, M. L., DePaolis, L. A., Snuggs, M. B., & Lee, P. (1991). Insulin inhibits transcription of the human gene for insulin-like growth factor-binding protein-1. Journal of Biological Chemistry, 266(28), 18868-18876.

Insulin inhibits transcription of the human gene for insulin-like growth factor-binding protein-1. / Powell, David R.; Suwanichkul, Adisak; Cubbage, Michael L.; DePaolis, Laura A.; Snuggs, Mark B.; Lee, Phillip.

In: Journal of Biological Chemistry, Vol. 266, No. 28, 05.10.1991, p. 18868-18876.

Research output: Contribution to journalArticle

Powell, DR, Suwanichkul, A, Cubbage, ML, DePaolis, LA, Snuggs, MB & Lee, P 1991, 'Insulin inhibits transcription of the human gene for insulin-like growth factor-binding protein-1', Journal of Biological Chemistry, vol. 266, no. 28, pp. 18868-18876.
Powell DR, Suwanichkul A, Cubbage ML, DePaolis LA, Snuggs MB, Lee P. Insulin inhibits transcription of the human gene for insulin-like growth factor-binding protein-1. Journal of Biological Chemistry. 1991 Oct 5;266(28):18868-18876.
Powell, David R. ; Suwanichkul, Adisak ; Cubbage, Michael L. ; DePaolis, Laura A. ; Snuggs, Mark B. ; Lee, Phillip. / Insulin inhibits transcription of the human gene for insulin-like growth factor-binding protein-1. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 28. pp. 18868-18876.
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