Chemical synthesis of the ubiquitinated form of histone H3 and its effect on DNA methyltransferase 1

Abstract

Posttranslational modifications of histone proteins, which form nucleosome cores, play an important role in gene regulation. Ubiquitination is one such modification. We previously reported on the synthesis of ubiquitinated histone H3 with an isopeptide mimetic structure. In this report, we describe the preparation of ubiquitinated histone H3 peptides with a native isopeptide structure, which showed a slightly weaker effect on the enzymatic activity of DNA methyltransferase 1 than the previous ubiquitinated H3 peptide analogs. These findings show that a native structure is important for determining the mechanism of the function, although ubiquitinated H3 peptide analogs can mimic the role of the original ubiquitinated H3. We also report on the successful preparation of the ubiquitinated full length histone H3.

Chemical synthesis of palmitoylated histone H4

“…1,2,3 To directly understand the function of a histone with specific modification(s), histones with modifications (methylation, acetylation, ubiquitination) on the specific target lysine residue were chemically synthesized. 4,5 By using synthetic histones, the functional interactions between the histone modifications and other epigenetic regulations, i.e., DNA modification, have been characterized. 6,7 Protein lipidation is also an important post-translational modification, and the modification has been reported to increase its affinity to membranes and subsequent interaction with other proteins for cell signaling.…”

Fmoc-Compatible and C-terminal-Sequence-Independent Peptide Alkyl Thioester Formation Using Cysteinylprolyl Imide

“…We applied the CPI-mediated DKP thioester preparation to the chemical synthesis of histone H3.2 − bearing K56Ac modification. H3K56Ac is incorporated to replicating DNA and enhances the ability of two histone chaperones CAF-1 and Rtt106 to assemble DNA into nucleosomes by increasing the binding affinity of these chaperones .…”

Fmoc-Compatible and External-Thiol-Free Peptide C-Terminus Alkyl Thioester Formation Using Cysteinylprolyl Imide

“…We applied the CPI-mediated DKP thioester preparation to the chemical synthesis of histone H3.2 [31][32][33][34][35][36][37][38][39] bearing K56Ac modification. H3K56Ac is incorporated to replicating DNA and enhances the ability of two histone chaperones CAF-1 and Rtt106 to assemble DNA into nucleosomes by increasing the binding affinity of these chaperones.…”