Gene expression in humans is controlled by the differential binding of ~1600 transcription factors (TF) to cis-regulatory elements throughout the genome. After binding to DNA, TF function is subsequently determined by interactions with a constellation of ~1100 cofactor proteins (COFs) that include chromatin-modifying enzymes, the Mediator complex, and other diverse regulators. These TF-COF interaction networks are cell state-specific, adapt in response to cell signaling events, and can be altered in disease. While much has been learned about TF-DNA binding from TFs from high-throughput biochemical and genomic assays, TF-COF networks in different cellular contexts remain poorly annotated and experimentally difficult to characterize. Affinity purification / Mass spectrometry (AP/MS)-based methods are currently the most widely used to characterize cell-specific protein-protein interactions, but these methods remain sufficiently labor intensive that rapid profiling of TF-COF networks for many factors and cellular conditions is prohibitive. To address this need, we have recently developed a protein-binding microarray (PBM)-based approach, called CoRec (Cofactor Recruitment), to rapidly profile DNA-bound TF-COF complexes in a cell-specific manner. I will discuss this method and its applications to problems of (1) defining TF-COF networks in cells, and (2) characterizing TF-COF complexes impacted by non-coding single-nucleotide polymorphisms (ncSNPs).

Rapid profiling of transcription factor-cofactor complexes in cells

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.