Alternative Splicing of G9a Regulates Neuronal Differentiation

Cell Rep. 2016 Mar 29;14(12):2797-808. doi: 10.1016/j.celrep.2016.02.063. Epub 2016 Mar 17.

Abstract

Chromatin modifications are critical for the establishment and maintenance of differentiation programs. G9a, the enzyme responsible for histone H3 lysine 9 dimethylation in mammalian euchromatin, exists as two isoforms with differential inclusion of exon 10 (E10) through alternative splicing. We find that the G9a methyltransferase is required for differentiation of the mouse neuronal cell line N2a and that E10 inclusion increases during neuronal differentiation of cultured cells, as well as in the developing mouse brain. Although E10 inclusion greatly stimulates overall H3K9me2 levels, it does not affect G9a catalytic activity. Instead, E10 increases G9a nuclear localization. We show that the G9a E10(+) isoform is necessary for neuron differentiation and regulates the alternative splicing pattern of its own pre-mRNA, enhancing E10 inclusion. Overall, our findings indicate that by regulating its own alternative splicing, G9a promotes neuron differentiation and creates a positive feedback loop that reinforces cellular commitment to differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing*
  • Animals
  • Azepines / pharmacology
  • Brain / metabolism
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Nucleus / metabolism
  • Exons
  • Fluorescence Resonance Energy Transfer
  • Genes, Reporter
  • HeLa Cells
  • Histone-Lysine N-Methyltransferase / antagonists & inhibitors
  • Histone-Lysine N-Methyltransferase / genetics*
  • Histone-Lysine N-Methyltransferase / metabolism
  • Histones / metabolism
  • Humans
  • Methylation / drug effects
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Neurons / cytology
  • Neurons / metabolism
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Quinazolines / pharmacology
  • RNA Interference
  • RNA Precursors / metabolism
  • RNA, Small Interfering / metabolism
  • Real-Time Polymerase Chain Reaction
  • Tretinoin / pharmacology

Substances

  • Azepines
  • BIX 01294
  • Histones
  • Protein Isoforms
  • Quinazolines
  • RNA Precursors
  • RNA, Small Interfering
  • Tretinoin
  • Histone-Lysine N-Methyltransferase