Split-luciferase Epigenetic Assays for Drug Discovery

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Project Summary Epigenetic control is essential for maintaining transcriptional integrity. Regulation of the dynamic epigenome is mediated through a range of post translational modifications on DNA and histone tails. Histone modifications are mediated by enzymes which can be broadly classified into “writers” that catalyze the addition of chemical modifications, “erasers” that remove the modifications, and “readers” that can recognize chemical modifications through specific protein domains. The writing and erasing of histone marks by enzymes is a dynamic process, and when dysregulated is associated with cancer, inflammatory and neurological diseases. In this application we focus on the two classes of histone modifying enzymes, lysine methyl transferases (KMTs) and lysine demethylases (KDMs), and aim to develop and validate cell- based assays targeting both KMTs and KDMs. These assays, based on a three-hybrid split luciferase system, are reversible allowing dose dependent quantification of inhibitor binding. These efforts are both significant and innovative as they can report on the direct binding of a drug to the target protein at its intended site of action, thereby facilitating the generation of lead therapeutic candidates and identification of chemical probes for studying histone biology and pathways.