Protein acetylation is one of the most frequent post-translational modifications (PTMs) in the human proteome and regulates myriad cellular processes. Dysregulation of protein acetylation is implicated in a broad range of diseases, such as cancer and immunological disorders. Despite its frequency and profound effects on protein function, there are limited research tools to dynamically regulate protein-specific acetylation in situ. Here, we developed a generalizable acetylation tagging system, called AceTAG, to induce acetylation of targeted proteins through the use of heterobifunctional small molecules. AceTAG molecules are designed to mediate ternary complex formation between an FKBP12^F36V tagged protein and the acetyltransferase p300/CBP. We show that these molecules are capable of inducing acetylation of various proteins in a dose-dependent fashion, including histone H3.3, NF-kB subunit p65/RelA, and the tumor suppressor p53. We further demonstrate that AceTAG-induced acetylation is rapid, reversible, and selective. Moreover, using p53 as a case study, we provide preliminary evidence that AceTAG can be used to investigate functional consequences of targeted acetylation directly in cells. Toward this end, we envision our AceTAG strategy as a promising method to investigate various roles of protein acetylation in cells. Looking forward, we predict that AceTAG will enable explorations of chemically-induced protein acetylation as a therapeutic strategy.

Schematic depiction of the AceTAG system. AceTAG molecules, composed of an FKBP12<sup>F36V</sup> ligand and a p300 bromodomain ligand, are designed to mediate chemically-induced acetylation of targeted protein.

Schematic depiction of the AceTAG system. AceTAG molecules, composed of an FKBP12^F36V ligand and a p300 bromodomain ligand, are designed to mediate chemically-induced acetylation of targeted protein.

Christopher Parker

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