β-actin regulates a heterochromatin landscape essential for optimal induction of neuronal programs during direct reprograming

PLoS Genet. 2018 Dec 17;14(12):e1007846. doi: 10.1371/journal.pgen.1007846. eCollection 2018 Dec.

Abstract

During neuronal development, β-actin serves an important role in growth cone mediated axon guidance. Consistent with this notion, in vivo ablation of the β-actin gene leads to abnormalities in the nervous system. However, whether β-actin is involved in the regulation of neuronal gene programs is not known. In this study, we directly reprogramed β-actin+/+ WT, β-actin+/- HET and β-actin-/- KO mouse embryonic fibroblast (MEFs) into chemically induced neurons (CiNeurons). Using RNA-seq analysis, we profiled the transcriptome changes among the CiNeurons. We discovered that induction of neuronal gene programs was impaired in KO CiNeurons in comparison to WT ones, whereas HET CiNeurons showed an intermediate levels of induction. ChIP-seq analysis of heterochromatin markers demonstrated that the impaired expression of neuronal gene programs correlated with the elevated H3K9 and H3K27 methylation levels at gene loci in β-actin deficient MEFs, which is linked to the loss of chromatin association of the BAF complex ATPase subunit Brg1. Together, our study shows that heterochromatin alteration in β-actin null MEFs impedes the induction of neuronal gene programs during direct reprograming. These findings are in line with the notion that H3K9Me3-based heterochromatin forms a major epigenetic barrier during cell fate change.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / deficiency
  • Actins / genetics
  • Actins / metabolism*
  • Animals
  • Cells, Cultured
  • Cellular Reprogramming / genetics
  • Epigenesis, Genetic
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Gene Knockout Techniques
  • Heterochromatin / genetics
  • Heterochromatin / metabolism*
  • Mice
  • Neurons / cytology*
  • Neurons / metabolism*

Substances

  • Actins
  • Heterochromatin

Grants and funding

The research was funded by grants from the Swedish Research Council (www.vr.se), Cancerfonden (www.cancerfonden.se) and New York University Abu Dhabi (nyuad.nyu.edu) to PP. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.