The expedient, CAET-assisted synthesis of dual-monoubiquitinated histone H3 enables evaluation of its interaction with DNMT1

Site-selective conjugation chemistry has proven effective to synthesize homogenously ubiquitinated histones. Recently, a powerful strategy using 2-((2-chloroethyl) amino) ethane-1-thiol (CAET) as a bifunctional handle was developed to generate chemically stable ubiquitin chains without racemization and homodimerization. Herein, we extend this strategy to the expedient synthesis of ubiquitinated histones, exemplifying its utility to not only synthesize single-monoubiquitinated histones, but dual-monoubiquitinated histones as well. The synthetic histones enabled us to evaluate the binding of DNMT1 to ubiquitinated nucleosomes and map the hotspots of this interaction. Our work highlights the potential of modern chemical protein synthesis to synthesize ubiquitinated histones for epigenetic studies.

Automated summary

Site-selective conjugation chemistry using CAET as a bifunctional handle has been developed to synthesize chemically stable ubiquitin chains. This strategy has been extended to the synthesis of ubiquitinated histones, enabling the study of DNMT1 binding to ubiquitinated nucleosomes and mapping the interaction hotspots. Our work highlights the potential of modern chemical protein synthesis for epigenetic studies.