Rescuing the aberrant sex development of H3K9 demethylase Jmjd1a-deficient mice by modulating H3K9 methylation balance

Shunsuke Kuroki; Naoki Okashita; Shoko Baba; Ryo Maeda; Shingo Miyawaki; Masashi Yano; Miyoko Yamaguchi; Satsuki Kitano; Hitoshi Miyachi; Akihiro Itoh; Minoru Yoshida; Makoto Tachibana

doi:10.1371/journal.pgen.1007034

Rescuing the aberrant sex development of H3K9 demethylase Jmjd1a-deficient mice by modulating H3K9 methylation balance

PLoS Genet. 2017 Sep 26;13(9):e1007034. doi: 10.1371/journal.pgen.1007034. eCollection 2017 Sep.

Affiliations

¹ Division of Epigenome Dynamics, Institute of Advanced Medical Sciences, Tokushima University, Tokushima, Japan.
² Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Sakyo-ku, Kyoto, Japan.
³ Chemical Genomics Research Group, RIKEN Center for Sustainable Resource Science, Wako, Saitama, Japan.

Abstract

Histone H3 lysine 9 (H3K9) methylation is a hallmark of heterochromatin. H3K9 demethylation is crucial in mouse sex determination; The H3K9 demethylase Jmjd1a deficiency leads to increased H3K9 methylation at the Sry locus in embryonic gonads, thereby compromising Sry expression and causing male-to-female sex reversal. We hypothesized that the H3K9 methylation level at the Sry locus is finely tuned by the balance in activities between the H3K9 demethylase Jmjd1a and an unidentified H3K9 methyltransferase to ensure correct Sry expression. Here we identified the GLP/G9a H3K9 methyltransferase complex as the enzyme catalyzing H3K9 methylation at the Sry locus. Based on this finding, we tried to rescue the sex-reversal phenotype of Jmjd1a-deficient mice by modulating GLP/G9a complex activity. A heterozygous GLP mutation rescued the sex-reversal phenotype of Jmjd1a-deficient mice by restoring Sry expression. The administration of a chemical inhibitor of GLP/G9a enzyme into Jmjd1a-deficient embryos also successfully rescued sex reversal. Our study not only reveals the molecular mechanism underlying the tuning of Sry expression but also provides proof on the principle of therapeutic strategies based on the pharmacological modulation of epigenetic balance.

MeSH terms

Animals
Female
Gene Expression Regulation
Genetic Loci
Gonads / embryology
Gonads / metabolism
Histone-Lysine N-Methyltransferase / genetics
Histone-Lysine N-Methyltransferase / metabolism*
Histones / genetics
Histones / metabolism*
Jumonji Domain-Containing Histone Demethylases / deficiency
Jumonji Domain-Containing Histone Demethylases / genetics*
Jumonji Domain-Containing Histone Demethylases / metabolism
Male
Methylation
Mice
Mice, Inbred C57BL
Mice, Knockout
SOX9 Transcription Factor / genetics
SOX9 Transcription Factor / metabolism
Sequence Analysis, DNA
Sex-Determining Region Y Protein / genetics
Sex-Determining Region Y Protein / metabolism*
Sexual Development / genetics*

Substances

Histones
SOX9 Transcription Factor
Sex-Determining Region Y Protein
Sox9 protein, mouse
Sry protein, mouse
Jumonji Domain-Containing Histone Demethylases
Kdm3a protein, mouse
G9a protein, mouse
GLP protein, mouse
Histone-Lysine N-Methyltransferase

Grants and funding

This work was supported by KAKENHI from the Japan Society for the Promotion of Science (https://www.jsps.go.jp/) Grant Numbers 26250037 (MT), 16H01218 (MT), 16H01409 (MT), 17H06424 (MT), 16K21196 (SKu), and 16K18492 (NO); Funding Program for Next Generation World-Leading Researchers (http://www.cao.go.jp/) (MT); Takeda Science Foundation (http://www.takeda-sci.or.jp/) (MT); the NOVARTIS Foundation (http://japanfoundation.novartis.org/) (SKu); and a Promotion of Science Cooperative Research Grant of the Institute for Enzyme Research, Joint Usage/Research Center, Tokushima University (HM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.