Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter

K. Kou, D. W. Mittanck, C. Fu, P. Rotwein

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The actions of insulin-like growth factors I and II (IGF-I and -II) are modulated by interactions with one or more of a family of secreted IGF binding proteins (IGFBPs). IGFBP-5, the most conserved of the six known IGFBPs, is a 252-amino-acid protein that has been shown both to potentiate and inhibit IGF action. In previous studies, we have cloned and characterized the mouse IGFBP-5 gene and demonstrated that it is expressed in a hierarchical pattern in different adult mouse tissues and during rodent embryonic development. In this report, we describe the initial analysis of the IGFBP-5 gene promoter. By transient gene transfer studies, we show the orientation-specific activity of DNA fragments containing from 31 to 4,100 bp from the 5'-flanking region of the mouse IGFBP-5 gene in directing expression of the heterologous reporter gene luciferase in Hep G2 cells. DNA fragments with only 156 bp of 5'-flanking sequence mediated over 60% of maximal promoter activity, and a segment containing the TATA box and the first 120 bp of exon 1 still conferred some promoter function. Within the highly active 156-bp region, we identified a 37-bp segment from -70 to -34 that exhibited specific binding in DNase I footprinting and gel-mobility shift experiments with Hep G2 nuclear protein extracts. The footprinted region, which is almost completely conserved in the rat and human IGFBP-5 genes, was responsible for at least 70% of the activity of the intact promoter, as evidenced by the deleterious consequences of small internal deletions within this sequence on promoter function. Taken together, our results demonstrate that the mouse IGFBP-5 gene promoter has a simple structure, and defines a framework for examining the mechanisms responsible for the multifactorial regulation of IGFBP-5 gene transcription during development and in the adult.

Original languageEnglish (US)
Pages (from-to)241-249
Number of pages9
JournalDNA and Cell Biology
Volume14
Issue number3
StatePublished - 1995
Externally publishedYes

Fingerprint

Insulin-Like Growth Factor Binding Protein 5
Genes
Insulin-Like Growth Factor Binding Proteins
Insulin-Like Growth Factor II
5' Flanking Region
Insulin-Like Growth Factor I
TATA Box
Sequence Deletion
Deoxyribonuclease I
DNA
Hep G2 Cells
Nuclear Proteins
mouse IGFBP5-interacting protein
Luciferases
Reporter Genes
Embryonic Development
Exons
Rodentia
Gels
Amino Acids

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Kou, K., Mittanck, D. W., Fu, C., & Rotwein, P. (1995). Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter. DNA and Cell Biology, 14(3), 241-249.

Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter. / Kou, K.; Mittanck, D. W.; Fu, C.; Rotwein, P.

In: DNA and Cell Biology, Vol. 14, No. 3, 1995, p. 241-249.

Research output: Contribution to journalArticle

Kou, K, Mittanck, DW, Fu, C & Rotwein, P 1995, 'Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter', DNA and Cell Biology, vol. 14, no. 3, pp. 241-249.
Kou, K. ; Mittanck, D. W. ; Fu, C. ; Rotwein, P. / Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter. In: DNA and Cell Biology. 1995 ; Vol. 14, No. 3. pp. 241-249.
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