Analysis of plant regulatory DNA sequences by the yeast-one-hybrid assay

Abstract

Regulatory DNA sequences harbor the essential information to control specific gene expression changes and integrate information derived from upstream signaling cascades. This regulatory potential is mediated by direct binding of proteins, e.g., transcription factors, to defined stretches of DNA motifs in regulatory regions. The analysis of these DNA regions, at which several signaling pathways could merge to orchestrate gene expression, is still a challenging task. To date, the combination of functional approaches in the laboratory and computer aided sequence evaluation is frequently used for regulatory sequence analysis. The yeast-one-hybrid method is a possible approach to test for direct binding of plant proteins to DNA in a heterologous system. Moreover, it is the most frequently used method for the identification of DNA-binding proteins targeting a given DNA sequence by screening a cDNA library.