title: Dual Genetic Encoding of Acetyl-lysine and Non-deacetylatable Thioacetyl-lysine Mediated by Flexizyme.
abstract: Acetylation of lysine residues is an important post-translational protein modification. Lysine acetylation in histones and its crosstalk with other post-translational modifications in histone and non-histone proteins are crucial to DNA replication, DNA repair, and transcriptional regulation. We incorporated acetyl-lysine (AcK) and the non-hydrolyzable thioacetyl-lysine (ThioAcK) into full-length proteins in vitro, mediated by flexizyme. ThioAcK and AcK were site-specifically incorporated at different lysine positions into human histone H3, either individually or in pairs. We demonstrate that the thioacetyl group in histone H3 could not be removed by the histone deacetylase sirtuin type 1. This method provides a powerful tool to study protein acetylation and its role in crosstalk between post-translational modifications.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, Whitney Avenue 266, New Haven, CT, 06511, USA.
authors.forename: Hai
authors.initials: H
authors.lastname: Xiong
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, Whitney Avenue 266, New Haven, CT, 06511, USA.
authors.forename: Noah M
authors.initials: NM
authors.lastname: Reynolds
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, Whitney Avenue 266, New Haven, CT, 06511, USA.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Fan
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, Whitney Avenue 266, New Haven, CT, 06511, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Englert
authors.affiliation: Yale Center for Molecular Discovery, Yale University, West Haven, CT, 06516, USA.
authors.forename: Denton
authors.initials: D
authors.lastname: Hoyer
authors.affiliation: Department of Chemistry, Yale University, 225 Prospect Street, New Haven, CT, 06511, USA.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Miller
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, Whitney Avenue 266, New Haven, CT, 06511, USA. dieter.soll@yale.edu.|Department of Chemistry, Yale University, 225 Prospect Street, New Haven, CT, 06511, USA. dieter.soll@yale.edu.
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
mesh_terms: D000107:Acetylation; D004260:DNA Repair; D004261:DNA Replication; D004798:Enzymes / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
chemical_list: D004798:Enzymes; D008239:Lysine
keywords: flexizyme; histone; lysine acetylation; post-translational modifications; thioacetyl-lysine