The interaction between a protein and DNA is involved in almost all cellular functions, and is vitally important in transcriptional regulation. There are two complementary approaches used to detect the interactions between a transcription factor (TF) and DNA, i.e., the TF-centered or protein-DNA approach, and the gene-centered or DNA-protein approach. The yeast one-hybrid (Y1H) is a powerful and widely used gene-centered system to identify DNA-protein interactions. However, a powerful and simple TF-centered method to study protein-DNA interactions like Y1H is lacking. Here, we provide a TF-centered method based on the Y1H system to identify the motifs recognized by a defined TF, termed TF-centered Y1H. In this system, a random short DNA sequence insertion library is generated as the prey DNA sequences to interact with a defined TF as the bait. TF-centered Y1H could identify quickly the motifs bound by a defined TF, representing a reliable and efficient approach with the advantages of Y1H. Therefore, this TF-centered Y1H may have a wide application in protein-DNA interaction studies.
Keywords: Cis-acting element; Gene expression regulation; Protein–DNA interaction; Transcription factor; Yeast one-hybrid.