A methylation-phosphorylation switch controls EZH2 stability and hematopoiesis

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Abstract

The Polycomb Repressive Complex 2 (PRC2) methylates H3K27 to regulate development and cell fate by transcriptional silencing. Alteration of PRC2 is associated with various cancers. Here, we show that mouse Kdm1a deletion causes dramatic reduction of PRC2 proteins, whereas mouse null mutation of L3mbtl3 or Dcaf5 results in PRC2 accumulation and increased H3K27 trimethylation. The catalytic subunit of PRC2, EZH2, is methylated at lysine 20 (K20), promoting EZH2 proteolysis by L3MBTL3 and the CLR4^DCAF5ubiquitin ligase. KDM1A (LSD1) demethylates the methylated K20 to stabilize EZH2. K20 methylation is inhibited by AKT-mediated phosphorylation of serine 21 in EZH2. Mouse Ezh2^K20R/K20R mutants develop hepatosplenomegaly associated with high GFI1B expression, and Ezh2^K20R/K20R mutant bone marrows expand hematopoietic stem cells and downstream hematopoietic populations. Our studies reveal that EZH2 is regulated by methylation-dependent proteolysis, which is negatively controlled by AKT-mediated S21 phosphorylation to establish a methylation-phosphorylation switch to control the PRC2 activity and hematopoiesis.

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All data generated 801 during this study are included in the manuscript. Uncropped immunoblots, immunostaining, and gel blot images are accessible as source data

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Pengfei Guo

Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Las Vegas, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-6734-6549
Rebecca C. Lim

Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Las Vegas, United States

Competing interests
The authors declare that no competing interests exist.
Keshari Rajawasam

Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Las Vegas, United States

Competing interests
The authors declare that no competing interests exist.
Tiffany Trinh

Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Las Vegas, United States

Competing interests
The authors declare that no competing interests exist.
Hong Sun

Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Las Vegas, United States

For correspondence
hong.sun@unlv.edu

Competing interests
The authors declare that no competing interests exist.
Hui Zhang

Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Las Vegas, United States

For correspondence
hui.zhang@unlv.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-6028-2554

Funding

National Institutes of Health (R15CA254827)

Hong Sun

National Institutes of Health (R01GM140185)

Hui Zhang

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments including breeding, housing, genotyping, and sample collection were conducted in accordance with the animal protocols approved by the institutional Animal Use and Care Committee (IACUC) and complied with all relevant ethical regulations at University of Nevada, Las Vegas. All procedures were conducted according to the National Institutes of Health (NIH) Guide for Care and Use of Laboratory Animals. The UNLV IACUC is an AAALAC approved facility and meets the NIH Guide for the Care and Use of Animals. protocols (IACUC-01161)711621 and (IACUC-01177)832146 described previously (Guo et al., 2022).

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.