Interaction of the CNC-bZIP factor TCF11/LCR-F1/Nrf1 with MafG: Binding-site selection and regulation of transcription

• Published in: Nucleic acids research 1998-01, Vol.26 (2), p.512-520
• Main Authors: Johnsen, Øyvind, Murphy, Paula, Prydz, Hans, Kolstø, Anne-Brit
• Format: Article
• Language: English
• Subjects: Base Sequence; Binding Sites; Carrier Proteins - genetics; Consensus Sequence; Dimerization; DNA - chemistry; DNA-Binding Proteins - metabolism; DNA-Binding Proteins - pharmacology; Erythroid-Specific DNA-Binding Factors; Gene Expression Regulation; Humans; Leucine Zippers; MafG Transcription Factor; Maltose-Binding Proteins; NF-E2 Transcription Factor; NF-E2 Transcription Factor, p45 Subunit; NF-E2-Related Factor 1; Recombinant Fusion Proteins; Repressor Proteins - metabolism; Repressor Proteins - pharmacology; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription, Genetic; Transfection
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/26.2.512

We have previously shown that the widely expressed human transcription factor TCF11/LCR-F1/Nrf1 interacts with small Maf proteins and binds to a subclass of AP1-sites. Such sites are required for β-globin 5′ DNase I hypersensitive site 2 enhancer activity, erythroid porphobilinogen deaminase inducibility, hemin responsiveness by heme-oxygenase 1 and expression of the gene NAD(P)H:quinone oxidoreductase1. Here we report the optimal DNA-binding sequences for TCF11/LCR-F1/Nrf1 alone and as a heterodimer with MafG, identified by using binding-site selection. The heterodimer recognises a 5′-TGCTgaGTCAT-3′ binding-site that is identical to the established NF-E2-site, the antioxidant response element and the heme-responsive element while the binding specificity of the homomer is less stringent. To investigate the activity of TCF11 through this selected site, both alone and in the presence of MafG, we have used a transient transfection assay. TCF11 alone activates transcription while MafG alone acts as a repressor. When co-expressed, MafG interferes with TCF11 transactivation in a dose dependent manner. This indicates that MafG protein, which heterodimerises efficiently with TCF11 in vitro (the heterodimer having a higher affinity for DNA than TCF11 alone), does not co-operate with TCF11 in transactivating transcription. We propose that since both these factors are widely expressed, they may act together to contribute to the negative regulation of this specific target site. Efficient positive regulation by TCF11 may require alternative partners with perhaps more restricted expression patterns.