Functional Investigations of p53 Acetylation Enabled by Heterobifunctional Molecules

ACS Chem Biol. 2024 Sep 20;19(9):1918-1929. doi: 10.1021/acschembio.4c00438. Epub 2024 Sep 9.

Abstract

Post-translational modifications (PTMs) dynamically regulate the critical stress response and tumor suppressive functions of p53. Among these, acetylation events mediated by multiple acetyltransferases lead to differential target gene activation and subsequent cell fate. However, our understanding of these events is incomplete due to, in part, the inability to selectively and dynamically control p53 acetylation. We recently developed a heterobifunctional small molecule system, AceTAG, to direct the acetyltransferase p300/CBP for targeted protein acetylation in cells. Here, we expand AceTAG to leverage the acetyltransferase PCAF/GCN5 and apply these tools to investigate the functional consequences of targeted p53 acetylation in human cancer cells. We demonstrate that the recruitment of p300/CBP or PCAF/GCN5 to p53 results in distinct acetylation events and differentiated transcriptional activities. Further, we show that chemically induced acetylation of multiple hotspot p53 mutants results in increased stabilization and enhancement of transcriptional activity. Collectively, these studies demonstrate the utility of AceTAG for functional investigations of protein acetylation.

MeSH terms

  • Acetylation
  • Cell Line, Tumor
  • Humans
  • Protein Processing, Post-Translational*
  • Tumor Suppressor Protein p53* / metabolism
  • p300-CBP Transcription Factors* / metabolism

Substances

  • Tumor Suppressor Protein p53
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor
  • TP53 protein, human