Novel genetic tools for probing individual H3 molecules in each nucleosome.

Abstract:

:In eukaryotes, genomic DNA is packaged into the nucleus together with histone proteins, forming chromatin. The fundamental repeating unit of chromatin is the nucleosome, a naturally symmetric structure that wraps DNA and is the substrate for numerous regulatory post-translational modifications. However, the biological significance of nucleosomal symmetry until recently had been unexplored. To investigate this issue, we developed an obligate pair of histone H3 heterodimers, a novel genetic tool that allowed us to modulate modification sites on individual H3 molecules within nucleosomes in vivo. We used these constructs for molecular genetic studies, for example demonstrating that H3K36 methylation on a single H3 molecule per nucleosome in vivo is sufficient to restrain cryptic transcription. We also used asymmetric nucleosomes for mass spectrometric analysis of dependency relationships among histone modifications. Furthermore, we extended this system to the centromeric H3 isoform (Cse4/CENP-A), gaining insights into centromeric nucleosomal symmetry and structure. In this review, we summarize our findings and discuss the utility of this novel approach.

journal_name

Curr Genet

journal_title

Current genetics

authors

Ichikawa Y,Kaufman PD

doi

10.1007/s00294-018-0910-0

subject

Has Abstract

pub_date

2019-04-01 00:00:00

pages

371-377

issue

2

eissn

0172-8083

issn

1432-0983

pii

10.1007/s00294-018-0910-0

journal_volume

65

pub_type

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