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Methyltransferase G9A Regulates Osteogenesis via Twist Gene Repression.

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  • 11 Department of Oral Health Sciences, Life Sciences Institute, Faculty of Dentistry, The Biomedical Research Centre, University of British Columbia, Vancouver, Canada.

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|June 24, 2017
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Summary

The histone methyltransferase G9A epigenetically regulates cranial neural crest cell differentiation timing. Its deletion delays ossification and reduces jaw size by impacting Twist gene expression and proliferation.

Keywords:
H3K9me2RUNX2Wnt1-Crecraniofacialepigeneticsintramembranous ossification

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Area of Science:

  • Developmental Biology
  • Epigenetics
  • Craniofacial Development

Background:

  • Cranial neural crest cells are crucial for craniofacial development.
  • Epigenetic modifications play a role in regulating cell differentiation timing.
  • Histone methylation, specifically H3K9me2, is a key epigenetic mark.

Purpose of the Study:

  • To investigate the role of histone H3 lysine 9 methyltransferase G9A in cranial neural crest cell differentiation.
  • To elucidate the epigenetic mechanisms controlling craniofacial skeletal development.
  • To understand the regulation of Twist genes by G9A during ossification.

Main Methods:

  • Conditional deletion of G9A in neural crest cells using Wnt1-Cre.
  • Phenotypic analysis of homozygous-null animals, including skeletal measurements.
  • Gene expression analysis of osteogenic and mesenchymal markers (RUNX2, TWIST, osteopontin).
  • In vitro studies using G9A inhibitor (BIX-01294) on facial micromass cultures.
  • Chromatin immunoprecipitation (ChIP) to assess H3K9me2 marks on Twist gene regulatory regions.

Main Results:

  • G9A deletion resulted in viable but smaller animals with incomplete ossification and shorter jaws.
  • Delayed expression of osteopontin and persistent TWIST expression in osteogenic mesenchyme of mutants.
  • G9A inhibition in vitro upregulated TWIST and decreased osteogenic markers.
  • G9A catalyzes repressive H3K9me2 marks on Twist1 and Twist2 regulatory regions, which are removed upon G9A inhibition.

Conclusions:

  • G9A and H3K9me2 are critical epigenetic regulators of Twist gene expression during craniofacial development.
  • Epigenetic control by G9A is essential for the temporal regulation of cranial neural crest cell differentiation and ossification.
  • This study provides novel insights into the role of epigenetic mechanisms in controlling tissue-specific gene expression during development.