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Live-Cell Epigenome Manipulation by Synthetic Histone Acylation Catalyst System

preprint
submitted on 24.04.2020 and posted on 27.04.2020 by Yusuke Fujiwara, Yuki Yamanashi, Yuko Sato, Tomoya Kujirai, Hitoshi Kurumizaka, Hiroshi Kimura, Kenzo Yamatsugu, Shigehiro Kawashima, Motomu Kanai

Chemical modifications of histones play a pivotal role in the epigenome and regulation of gene expression, and their abnormality is tightly linked to numerous disease states in humans. Therefore, chemical tools to manipulate epigenome hold promise for both therapy and the elucidation of epigenetic mechanisms. We previously developed the chemical catalyst LANA-DSH, which binds to nucleosomes via a LANA peptide ligand, and selectively acylates proximal histone H2BK120 to the catalyst moiety by acti- vating acyl-CoAs. Thus far, however, histone acylation by a chem- ical catalyst system in living cells has not yet been demonstrated. Here, we report a chemical catalyst system, composed of a nucleo- some-binding catalyst (PEG-LANA-DSH) and a cell-permeable thioester acyl donor (NAC-acyl), that can promote regioselective lysine acylation of histones in living cells. Whereas LANA-DSH is rapidly decomposed in cells, attachment of polyethylene glycol (PEG) to the LANA moiety can prevent this undesired degradation. Increasing the size of PEG conferred LANA with greater in-cell stability, but reduced catalytic activity, indicating that there is an optimum PEG length balancing stability and catalytic activity. The optimized PEG-LANA-DSH catalyst 11 efficiently promoted H2BK120 acetylation in living cells, which subsequently sup- pressed ubiquitination of H2BK120, a mark regulating various chromatin functions, such as transcription and DNA damage re- sponse. Thus, our chemical catalyst system will be useful as a unique tool to manipulate the epigenome for therapeutic purposes or further understanding epigenetic mechanisms.

Funding

JP17H01522

JP17K19479

JP20H00489

JP19KK0179

JP18H04536

JP18H05534

JP17H01417

JP18H05527

JP20am0101076

JPMJER1901

History

Email Address of Submitting Author

skawashima@mol.f.u-tokyo.ac.jp

Institution

The University of Tokyo

Country

Japan

ORCID For Submitting Author

0000-0002-2946-6627

Declaration of Conflict of Interest

no conflict of interest

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