Targeted Protein Acetylation in Cells Using Heterobifunctional Molecules

Abstract

Protein acetylation is a central event in orchestrating diverse cellular processes. However, current strategies to investigate protein acetylation in cells are often nonspecific or lack temporal and magnitude control. Here, we developed an acetylation tagging system, AceTAG, to induce acetylation of targeted proteins. The AceTAG system utilizes bifunctional molecules to direct the lysine acetyltransferase p300/CBP to proteins fused with the small protein tag FKBP12F36V, resulting in their induced acetylation. Using AceTAG, we induced targeted acetylation of a diverse array of proteins in cells, specifically histone H3.3, the NF-κB subunit p65/RelA, and the tumor suppressor p53. We demonstrate that targeted acetylation with the AceTAG system is rapid, selective, reversible and can be controlled in a dose-dependent fashion. AceTAG represents a useful strategy to modulate protein acetylation and should enable the exploration of targeted acetylation in basic biological and therapeutic contexts.

Chasing molecular glue degraders: screening approaches

“…To conclude, the advances in the TPD field have fueled interest in other proximity-inducing concepts whose evolution will be interesting to follow in the coming years. [185][186][187][188][189] As our understanding of the molecular features that govern drug-induced productive PPIs grows and the use of this pharmacology in disease matures, new breakthroughs are sure to follow.…”

Methods to characterize and discover molecular degraders in cells

“…18B), acetylation-tagging system (AceTAG) to recruit acetylation (Fig. 18C), 138 and nanobody-engineered O -GlcNAc transferase (OGT) 139 or split O -GlcNAcase (OGA) 140 for targeted protein glycosylation and deglycosylation (Fig. 18D).…”

Beyond Proteolysis-Targeting Chimeric Molecules: Designing Heterobifunctional Molecules Based on Functional Effectors

“…Recently, novel heterobifunctional molecules that tether various effectors to a variety of targets have been developed . By recruiting RNases to specific transcripts to degrade RNAs, ribonuclease-targeting chimeras (RIBOTACs) were proposed in 2018. , Autophagy-targeting chimera (AUTAC) and lysosome-targeting chimera (LYTAC) technology, developed in 2019 and 2020, respectively, have been reported to work through the endosome/autophagosome pathway to degrade POIs. − In addition to degradation, heterobifunctional molecules such as phosphorylation-inducing chimeric small molecules (PHICS), protein phosphatase-recruiting chimeras (PHORCs/PhosTACs), and acetyltransferase-recruiting chimeras (AceTAGs) have been suggested to be PTM modulators. − These heterobifunctional molecules tether a kinase, protein phosphatase, or acetyltransferase to a POI, inducing the phosphorylation, dephosphorylation, or acetylation of the POI, respectively. With an increasing number of heterobifunctional molecules being reported, a strategy based on specific binding of proteins to bring them into proximity is the basis of a new generation of therapeutics and is providing new opportunities for additional small-molecule design …”