| pubmed-article:8288615 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C1416657 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C0012854 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C0005283 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C0040648 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C1704675 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C0596988 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C0215398 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C1521761 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C0936012 | lld:lifeskim |
| pubmed-article:8288615 | lifeskim:mentions | umls-concept:C1516960 | lld:lifeskim |
| pubmed-article:8288615 | pubmed:issue | 2 | lld:pubmed |
| pubmed-article:8288615 | pubmed:dateCreated | 1994-2-22 | lld:pubmed |
| pubmed-article:8288615 | pubmed:abstractText | We describe functional tests and molecular modeling of erythroid Krüppel-like factor (EKLF) interactions with its DNA binding site. EKLF, a zinc finger-containing, erythroid-specific transcription factor, binds and transactivates from the CACCC element, an evolutionarily conserved DNA sequence present within a large number of erythroid-specific promoters and enhancers. This DNA binding element is the site of naturally occurring point mutations that give rise to beta-thalassemia. We have directly tested whether CAC site point mutations (including two of the beta-thalassemia mutants) affect EKLF transactivation and DNA binding function. In vivo analyses demonstrate that EKLF is unable to transactivate a reporter plasmid that contains these mutations. In vitro analyses reveal a 40-100-fold decrease in binding affinity for these sites that accounts for the in vivo observations. The homology between the three EKLF and Zif268 zinc fingers and their conserved sequence-specific contacts to their target site allowed us to formulate a molecular model of the EKLF/CAC site complex, based primarily on energy minimization/refinement of the Zif268/DNA co-crystal structure. These models suggest that both specific and nonspecific hydrogen bonding play a critical role in the ability of EKLF to prefer binding to its cognate site. Analysis of sequence-specific contacts by EKLF to its target site within the beta-globin promoter verified the residues predicted to be important by the functional and modeling data. Together these results demonstrate that EKLF displays a strong discriminatory ability among potential DNA target sites consistent with the beta-thalassemia data. They also suggest that lack of EKLF binding to these sites may play a determining role in its phenotype, and they strengthen the evidence in favor of EKLF's proposed role in erythroid-specific transcriptional activation through the CACCC elements. | lld:pubmed |
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| pubmed-article:8288615 | pubmed:language | eng | lld:pubmed |
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| pubmed-article:8288615 | pubmed:citationSubset | IM | lld:pubmed |
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| pubmed-article:8288615 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:8288615 | pubmed:month | Jan | lld:pubmed |
| pubmed-article:8288615 | pubmed:issn | 0021-9258 | lld:pubmed |
| pubmed-article:8288615 | pubmed:author | pubmed-author:FengW CWC | lld:pubmed |
| pubmed-article:8288615 | pubmed:author | pubmed-author:BiekerJ JJJ | lld:pubmed |
| pubmed-article:8288615 | pubmed:author | pubmed-author:SouthwoodC... | lld:pubmed |
| pubmed-article:8288615 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:8288615 | pubmed:day | 14 | lld:pubmed |
| pubmed-article:8288615 | pubmed:volume | 269 | lld:pubmed |
| pubmed-article:8288615 | pubmed:geneSymbol | EKLF | lld:pubmed |
| pubmed-article:8288615 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:8288615 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:8288615 | pubmed:pagination | 1493-500 | lld:pubmed |
| pubmed-article:8288615 | pubmed:dateRevised | 2008-11-21 | lld:pubmed |
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| pubmed-article:8288615 | pubmed:year | 1994 | lld:pubmed |
| pubmed-article:8288615 | pubmed:articleTitle | Analyses of beta-thalassemia mutant DNA interactions with erythroid Krüppel-like factor (EKLF), an erythroid cell-specific transcription factor. | lld:pubmed |
| pubmed-article:8288615 | pubmed:affiliation | Department of Physiology, Mount Sinai School of Medicine, New York, New York 10029. | lld:pubmed |
| pubmed-article:8288615 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:8288615 | pubmed:publicationType | Comparative Study | lld:pubmed |
| pubmed-article:8288615 | pubmed:publicationType | Research Support, U.S. Gov't, P.H.S. | lld:pubmed |
| pubmed-article:8288615 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
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