ProTailor · Chemically Tailored Transcription Factor Proteins for Dissecting Gene Expression Regulation

The aberrant function of oncogenic transcription factors (TFs) is one of the most common drivers in human cancers, yet the mechanisms underlying their deregulation and targeting remain elusive. Understanding pathways that regulate TFs function has the potential to develop novel therapeutic compounds to target tumorigenesis. The overreaching goal of ProTailor is to develop ground-breaking modified TFs to understand gene expression regulation mechanisms and set the foundation for therapeutic opportunities to study and target human tumorigenesis.Aim 1: I will develop novel synthetic strategies to prepare site-specifically modified oncogenic TF analogs bearing defined post-translational modifications (PTMs) to decipher the molecular role of PTMs in regulating gene expression. I will combine chemoselective peptide ligation, and orthogonal transformations, along with high-throughput screening to probe the molecular role of essential PTMs on gene recognition, interaction, and cancer-related oncogene expression.Aim 2: I will generate libraries of chromatinized-DNA probes to investigate the interactions of modified TFs with packed DNA, aiming to understand how PTMs modulate TF-DNA interactions, accessibility, and influence gene expression regulation. I will probe the crosstalk between TF-PTMs, epigenetic modifications, and chromatin binding sites to unravel the mechanisms underlying gene recognition and interactions.Aim 3: I will design photoreactive TF probes to map the regulatory role of PTMs in the interactions of oncogenic TF analogs with transcriptional machinery. I will deliver the synthetic TF probes to cancer cells and dissect the mechanisms by which PTMs modulate biomolecular interactions and unravel potential PTM markers associated with oncogene expression. ProTailor will provide novel and powerful research tools to understand gene expression regulation mechanisms and lay the groundwork for therapeutic strategies to investigate and target human tumorigenesis.