University of Massachusetts Medical School Faculty Publications

UMMS Affiliation

Department of Molecular, Cell, and Cancer Biology; Department of Biochemistry and Molecular Pharmacology

Publication Date

9-6-2017

Document Type

Article Preprint

Disciplines

Amino Acids, Peptides, and Proteins | Cells | Genetic Phenomena | Investigative Techniques | Molecular Biology

Abstract

The repeating subunit of chromatin, the nucleosome, includes two copies of each of the four core histones, and several recent studies have reported that asymmetrically modified nucleosomes occur at regulatory elements in vivo. To probe the mechanisms by which histone modifications are read out, we designed an obligate pair of H3 heterodimers, termed H3X and H3Y, which we validated genetically and biochemically. Comparing the effects of asymmetric histone tail point mutants with those of symmetric double mutants revealed that a single methylated H3K36 per nucleosome was sufficient to silence cryptic transcription in vivo. We also demonstrate the utility of this system for analysis of histone modification crosstalk, using mass spectrometry to separately identify modifications on both H3 molecules within asymmetric nucleosomes. The ability to generate asymmetric nucleosomes in vivo provides a powerful tool to probe the mechanism by which H3 tails are read out by effector proteins in the cell.

Keywords

molecular biology, nucleosome symmetry, histones, heterodimers, asymmetric nucleosomes

Rights and Permissions

The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.

DOI of Published Version

10.1101/170811

Source

bioRxiv 170811; doi: https://doi.org/10.1101/170811. Link to preprint on bioRxiv service.

Comments

Full author list omitted for brevity. For the full list of authors, see article.

Related Resources

Now published in eLife doi: 10.7554/eLife.28836.

Journal/Book/Conference Title

bioRxiv

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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