Student The Hebrew University of Jerusalem The Hebrew University of Jerusalem, Israel
Abstract: The tumor suppressor protein p53, mutated in half of human cancers, plays a crucial role in cellular responses to DNA damage and genome stability, making it a key target for advancing cancer treatment and diagnosis. To identify genes and pathways critical for resistance to p53 upregulation, we performed a genome-wide CRISPR-Cas9 loss-of-function screen using Nutlin-3a, an inhibitor of the p53-MDM2 interaction that induces p53 accumulation and apoptotic cell death. Our findings revealed three enriched pathways: heparan sulfate glycosaminoglycan biosynthesis, diphthamide biosynthesis, and the Hippo pathway, with our study further confirming the interaction between the p53 and Hippo pathways. Notably, we discovered that the E3 ubiquitin-protein ligase TRIP12 is essential for the transcriptional activation of specific pro-apoptotic p53 target genes, with RNA-seq analysis in TRIP12 knockout cell lines revealing selective effects on p53’s transcriptional activity and highlighting its role in modulating p53-dependent gene expression. By uncovering novel pathways and mechanisms regulating p53 function, our study expands the understanding of p53 biology and provides valuable insights that may guide the development of innovative therapeutic strategies for combating cancer.
