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Uncovering DNA binding properties of the GATA4 and TBX5 transcription factor complex
Author(s) -
RosadoRodríguez Emili Patricia,
RodríguezRíos Jessica,
RodríguezMartínez José
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.lb185
Subject(s) - gata4 , transcription factor , systematic evolution of ligands by exponential enrichment , dna , biology , electrophoretic mobility shift assay , computational biology , gene , dna binding protein , microbiology and biotechnology , genetics , rna
Transcription factors (TFs) are essential master regulators of gene expression. These DNA binding proteins are fundamental for cellular differentiation during development. Eukaryotic TFs are notorious for not working alone and they often bind DNA as multimeric protein complexes to regulate gene expression. GATA4 and TBX5 are transcription factors that are central components of the gene regulatory network of heart development and function. Recent studies have determined the binding specificity of multiple transcription factors as monomers. However, the DNA‐binding properties of the cooperative complex between GATA4 and TBX5 remain undetermined. Based on this, we want to know the intrinsic DNA binding preferences and the emergent properties that result from the cooperative complex formed by GATA4 and TBX5. We used a cell‐free protein expression system to produce GATA4 and TBX5, and we successfully validated their DNA‐binding activity through an Electrophoretic Mobility Shift Assay (EMSA). Subsequently, we will determine the in vitro DNA‐binding specificity of the GATA4:TBX5 complex using Cognate Site Identification (CSI) by High‐Throughput Systematic Evolution of Ligands by Exponential Enrichment (HT‐SELEX). The findings of this study will help understand the DNA‐recognition rules of the GATA4:TBX5 complex and its potential roles in normal heart development. Support or Funding Information Grant Acknowledgement: This research is funded by the National Institutes of Health ‐ NIH SC1GM127231 and the Puerto Rico Louis Stokes Alliance for Minority Participation (PR‐LSAMP)‐Undergraduate Fellowship. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .