A Modular Platform for Effector Discovery in Induced-Proximity Lysine Acetylation.

Area of Science:

• Cellular Biology
• Molecular Biology
• Biochemistry

Background:

• Post-translational modifications (PTMs) are crucial for cellular functions and disease development.
• Current methods for manipulating PTMs via induced proximity require extensive synthesis and validation of effector molecules.

Purpose of the Study:

• To develop a modular platform for rapid evaluation of PTM editing enzymes in living cells.
• To accelerate the identification of effective enzyme-substrate relationships for targeted PTM manipulation.

Main Methods:

• Utilized a modular platform employing compound-dependent or nanobody-mediated induced proximity.
• Tested the platform using lysine acetylation as a model system, recruiting various acetyltransferases.
• Demonstrated programmable acetylation of GFP, histone H3, and p53.

Main Results:

• Successfully demonstrated programmable and site-specific acetylation of target proteins.
• Showcased selective PTM deposition across different substrates and cellular compartments.
• Validated the platform's ability to rapidly identify productive effector-substrate interactions.

Conclusions:

• The developed platform enables rapid screening of PTM editing enzymes.
• Facilitates the design of induced-proximity chemical probes for targeted PTM editing.
• Significantly accelerates research in PTM regulation and its role in disease.