Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins

Post-translational modification of the histone proteins in chromatin plays a central role in epigenetic control of DNA-templated processes in eukaryotic cells. Developing methods that enable the structure of histones to be manipulated is therefore essential to understand the biochemical mechanisms u...

Full description

Bibliographic Details
Main Authors:	David, Yael, Vila-Perelló, Miquel, Verma, Shivam, Muir, Tom W.
Format:	Online
Language:	English
Published:	2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4617616/

id	pubmed-4617616
recordtype	oai_dc
spelling	pubmed-46176162015-11-01 Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins David, Yael Vila-Perelló, Miquel Verma, Shivam Muir, Tom W. Article Post-translational modification of the histone proteins in chromatin plays a central role in epigenetic control of DNA-templated processes in eukaryotic cells. Developing methods that enable the structure of histones to be manipulated is therefore essential to understand the biochemical mechanisms underlying genomic regulation. Here we present a synthetic biology method to engineer histones bearing site-specific modifications on cellular chromatin using protein trans-splicing. We genetically fused the N-terminal fragment of ultrafast split-intein to the C-terminus of histone H2B, which upon reaction with a complementary synthetic C-intein, generated labeled histone. Using this approach, we incorporated various non-native chemical modifications to chromatin in vivo with temporal control. Furthermore, the time and concentration dependence of protein trans-splicing performed in nucleo enabled us to examine differences in the accessibility of the euchromatin and heterochromatin regions of the epigenome. Finally, we used protein trans-splicing to semi-synthesize a native histone modification, H2BK120 ubiquitination, in isolated nuclei, and show that this can trigger downstream epigenetic cross-talk of H3K79 methylation. 2015-04-06 2015-05 /pmc/articles/PMC4617616/ /pubmed/25901817 http://dx.doi.org/10.1038/nchem.2224 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	David, Yael Vila-Perelló, Miquel Verma, Shivam Muir, Tom W.
spellingShingle	David, Yael Vila-Perelló, Miquel Verma, Shivam Muir, Tom W. Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
author_facet	David, Yael Vila-Perelló, Miquel Verma, Shivam Muir, Tom W.
author_sort	David, Yael
title	Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
title_short	Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
title_full	Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
title_fullStr	Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
title_full_unstemmed	Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
title_sort	chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins
description	Post-translational modification of the histone proteins in chromatin plays a central role in epigenetic control of DNA-templated processes in eukaryotic cells. Developing methods that enable the structure of histones to be manipulated is therefore essential to understand the biochemical mechanisms underlying genomic regulation. Here we present a synthetic biology method to engineer histones bearing site-specific modifications on cellular chromatin using protein trans-splicing. We genetically fused the N-terminal fragment of ultrafast split-intein to the C-terminus of histone H2B, which upon reaction with a complementary synthetic C-intein, generated labeled histone. Using this approach, we incorporated various non-native chemical modifications to chromatin in vivo with temporal control. Furthermore, the time and concentration dependence of protein trans-splicing performed in nucleo enabled us to examine differences in the accessibility of the euchromatin and heterochromatin regions of the epigenome. Finally, we used protein trans-splicing to semi-synthesize a native histone modification, H2BK120 ubiquitination, in isolated nuclei, and show that this can trigger downstream epigenetic cross-talk of H3K79 methylation.
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4617616/
_version_	1613491909083267072

Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins

Similar Items