Cdk8 is required for establishment of H3K27me3 and gene repression by Xist and mouse development

Development. 2020 Jun 11;147(11):dev175141. doi: 10.1242/dev.175141.

Abstract

We previously identified the cyclin dependent kinase Cdk8 as a putative silencing factor for Xist To investigate its role in X inactivation, we engineered a Cdk8 mutation in mouse embryonic stem cells (ESCs) carrying an inducible system for studying Xist function. We found that Xist repressed X-linked genes at half of the expression level in Cdk8 mutant cells, whereas they were almost completely silenced in the controls. Lack of Cdk8 impaired Ezh2 recruitment and the establishment of histone H3 lysine 27 tri-methylation but not PRC1 recruitment by Xist Transgenic expression of wild-type but not catalytically inactive Cdk8 restored efficient gene repression and PRC2 recruitment. Mutation of the paralogous kinase Cdk19 did not affect Xist function, and combined mutations of Cdk8 and Cdk19 resembled the Cdk8 mutation. In mice, a Cdk8 mutation caused post-implantation lethality. We observed that homozygous Cdk8 mutant female embryos showed a greater developmental delay than males on day 10.5. Together with the inefficient repression of X-linked genes in differentiating Cdk8 mutant female ESCs, these data show a requirement for Cdk8 in the initiation of X inactivation.

Keywords: Cdk8; Cyclin dependent kinase; Gene regulation; Polycomb; X inactivation; Xist.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cyclin-Dependent Kinase 8 / genetics
  • Cyclin-Dependent Kinase 8 / metabolism*
  • Cyclin-Dependent Kinases / genetics
  • Cyclin-Dependent Kinases / metabolism
  • Embryo, Mammalian
  • Embryonic Development
  • Enhancer of Zeste Homolog 2 Protein / metabolism
  • Female
  • Gene Editing
  • Gene Expression Regulation, Developmental
  • Histones / metabolism*
  • Male
  • Methylation
  • Mice
  • Mice, Transgenic
  • Mouse Embryonic Stem Cells / cytology
  • Mouse Embryonic Stem Cells / metabolism
  • Mutagenesis
  • Polycomb Repressive Complex 2 / metabolism
  • RNA, Guide, CRISPR-Cas Systems
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • SOXB1 Transcription Factors / deficiency
  • SOXB1 Transcription Factors / genetics

Substances

  • Histones
  • RNA, Long Noncoding
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • XIST non-coding RNA
  • Enhancer of Zeste Homolog 2 Protein
  • Ezh2 protein, mouse
  • Polycomb Repressive Complex 2
  • CDK19 protein, mouse
  • Cdk8 protein, mouse
  • Cyclin-Dependent Kinase 8
  • Cyclin-Dependent Kinases

Associated data

  • Dryad/10.5061/dryad.tqjq2bvw0