Document Detail


Identification of a key regulatory element for the basal activity of the human insulin-like growth factor II gene promoter P3.
MedLine Citation:
PMID:  9581544     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Transcription of the human insulin-like growth factor II (IGF-II) gene is under the control of four promoters (P1-P4) that are differentially active during growth and development. Promoter 3 (P3) is the most active promoter during fetal development as well as in most adult tissues. P3 is also the most active promoter in tumour tissues and cell lines expressing IGF-II. Transient transfections of HeLa and Hep3B cells with truncated promoter constructs revealed that the region between -289 and -183 relative to the transcription start site supports basal promoter activity in both cell lines. Footprint experiments showed that the region between positions -192 and -172 (P3-4) is the only element bound by nuclear proteins. P3-4 is bound by five proteins, of which three proteins (proteins 3, 4 and 5) bind specifically and are expressed at the same levels in HeLa and Hep3B cells. Electrophoretic mobility shift assays and differential footprint experiments revealed the presence of two protein-binding regions within the P3-4 element. Proteins 4 and 5 bind box A (-193 to -188), whereas box B (-183 to -172) is bound by protein 3. From transcription experiments in vitro it can be concluded that Box A is essential for P3 activity. Box A is part of a region 11 dG residues long and is protected by proteins 4 and 5 that bind a contiguous set of six dG residues. DNA-binding of proteins 4 and 5 to box A requires the presence of Zn2+ ions. Thus structural and functional analysis reveals that the P3-4 element is a key regulatory element of P3 that contains two separate binding sites for proteins essential for the basal activity of IGF-II P3.
Authors:
L E Rietveld; P E Holthuizen; J S Sussenbach
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  327 ( Pt 3)     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1997 Nov 
Date Detail:
Created Date:  1998-05-20     Completed Date:  1998-05-20     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  689-97     Citation Subset:  IM    
Affiliation:
Laboratory for Physiological Chemistry, Utrecht University, Stratenum, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Animals
DNA Footprinting
DNA-Binding Proteins / genetics*,  metabolism
Deoxyribonuclease I / metabolism
Drosophila Proteins
GATA Transcription Factors
Gene Expression Regulation*
Hela Cells
Humans
Insulin-Like Growth Factor II / genetics*,  metabolism
Mice
Promoter Regions, Genetic*
Transcription Factors / metabolism
Transcription, Genetic
Transfection
Zinc / metabolism
Chemical
Reg. No./Substance:
0/DNA-Binding Proteins; 0/Drosophila Proteins; 0/GATA Transcription Factors; 0/Transcription Factors; 0/serpent protein, Drosophila; 67763-97-7/Insulin-Like Growth Factor II; 7440-66-6/Zinc; EC 3.1.21.1/Deoxyribonuclease I
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