Accurate H3K27 methylation can be established de novo by SUZ12-directed PRC2

Nature Structural & Molecular Biology
Jonas W HøjfeldtKristian Helin

Abstract

Polycomb repressive complex 2 (PRC2) catalyzes methylation on lysine 27 of histone H3 (H3K27) and is required for maintaining transcriptional patterns and cellular identity, but the specification and maintenance of genomic PRC2 binding and H3K27 methylation patterns remain incompletely understood. Epigenetic mechanisms have been proposed, wherein pre-existing H3K27 methylation directs recruitment and regulates the catalytic activity of PRC2 to support its own maintenance. Here we investigate whether such mechanisms are required for specifying H3K27 methylation patterns in mouse embryonic stem cells (mESCs). Through re-expression of PRC2 subunits in PRC2-knockout cells that have lost all H3K27 methylation, we demonstrate that methylation patterns can be accurately established de novo. We find that regional methylation kinetics correlate with original methylation patterns even in their absence, and specification of the genomic PRC2 binding pattern is retained and specifically dependent on the PRC2 core subunit SUZ12. Thus, the H3K27 methylation patterns in mESCs are not dependent on self-autonomous epigenetic inheritance.

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Datasets Mentioned

BETA
GSE103685
PRJNA404057
SRP117246

Methods Mentioned

BETA
immunoprecipitation
genetic knockout
ChIP-seq
RNA-seq
gene trap
ChIP
FACS

Software Mentioned

Bowtie2
DESeq2
Easeq
Trimmomatic
bedtools
GenomicFeatures
STAR
Illuminaclip
Bioconducter

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