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Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones
Published on: March 26, 2017
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Examining histone modification crosstalk using immobilized libraries established from ligation-ready nucleosomes
Diego Aparicio Pelaz1, Zhadyra Yerkesh2, Sören Kirchgäßner1
1Interfaculty Institute of Biochemistry, University of Tübingen Auf der Morgenstelle 34 D-72076 Tübingen Germany dirk.schwarzer@uni-tuebingen.de.
Chemical Science
|June 14, 2021
Summary
Researchers developed a new method to create diverse histone modifications on nucleosomes, enabling the discovery of new chromatin regulatory mechanisms. This approach revealed a novel interaction between histone modifications and HP1 protein recruitment.
Area of Science:
- Molecular Biology
- Epigenetics
- Chromatin Biology
Background:
- Chromatin signaling involves numerous histone posttranslational modifications (PTMs).
- Understanding histone PTMs requires access to specifically modified chromatin substrates.
- Current methods for generating modified histones are often laborious and time-consuming.
Purpose of the Study:
- To develop an efficient strategy for generating libraries of differentially modified nucleosomes.
- To bypass the need for extensive individual synthesis and assembly of modified nucleosomes.
- To explore the functional impact of complex histone modification patterns on chromatin regulation.
Main Methods:
- Utilized ligation-ready, immobilized nucleosomes reconstituted in a single batch.
- Employed sortase-mediated ligation for histone H3 and intein splicing for histone H2A.
- Generated libraries of up to 280 individually modified nucleosomes in a 96-well plate format.
Main Results:
- Successfully generated libraries of differentially modified nucleosomes.
- Screening revealed a novel long-range cross-talk between histone modifications.
- H3S28 phosphorylation was found to enhance HP1 protein recruitment to H3K9 methylated nucleosomes.
Conclusions:
- Ligation-ready nucleosomes provide efficient access to complex histone modification patterns.
- This strategy facilitates the discovery and dissection of chromatin regulatory principles.
- The findings highlight the importance of nucleosomal context in mediating histone PTM functions.
