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Live-cell Imaging Reveals the Dynamics of PRC2 and Recruitment to Chromatin by SUZ12-associated Subunits

Live-cell Imaging Reveals the Dynamics of PRC2 and Recruitment to Chromatin by SUZ12-associated Subunits

Daniel T Youmans, Jens C Schmidt, Thomas R Cech

Genes Dev. 2018 Jun 1;32(11-12):794-805.

PMID: 29891558

Abstract:

Polycomb-repressive complex 2 (PRC2) is a histone methyltransferase that promotes epigenetic gene silencing, but the dynamics of its interactions with chromatin are largely unknown. Here we quantitatively measured the binding of PRC2 to chromatin in human cancer cells. Genome editing of a HaloTag into the endogenous EZH2 and SUZ12 loci and single-particle tracking revealed that ∼80% of PRC2 rapidly diffuses through the nucleus, while ∼20% is chromatin-bound. Short-term treatment with a small molecule inhibitor of the EED-H3K27me3 interaction had no immediate effect on the chromatin residence time of PRC2. In contrast, separation-of-function mutants of SUZ12, which still form the core PRC2 complex but cannot bind accessory proteins, revealed a major contribution of AEBP2 and PCL homolog proteins to chromatin binding. We therefore quantified the dynamics of this chromatin-modifying complex in living cells and separated the contributions of H3K27me3 histone marks and various PRC2 subunits to recruitment of PRC2 to chromatin.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42414888	AEBP2 human			Price

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