Abstract
TGFβ-Inducible Early Gene (TIEG) and the alternatively-transcribed Early Growth Response Gene α (EGRα) share a Cys2His2 three-zinc finger region with high homology to Sp1 within its zinc finger region. Three-zinc finger transcription factors bind to GC-rich sequences, with small variations in consensus sequence between subfamilies. In this work, a consensus sequence was identified for TIEG/EGRα by expressing and purifying the zinc finger region of the protein, and using this to select a binding site from a random oligonucleotide library by iterative cycles of nitrocellulose filter binding and PCR. A fusion of the TIEG/EGRα with the VP16 activation domain supported transcription from this site when cloned in front of a heterologous promoter. Mutational analysis of the binding site identified a GT-rich core (5′-GGTGTG-3′) that was necessary for binding, with mutations outside of this region causing only a small to moderate decrease in binding.
Original language | English (US) |
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Pages (from-to) | 187-199 |
Number of pages | 13 |
Journal | DNA and Cell Biology |
Volume | 22 |
Issue number | 3 |
DOIs | |
State | Published - Mar 1 2003 |
ASJC Scopus subject areas
- Molecular Biology
- Genetics
- Cell Biology