TEAD transcription factors are required for normal primary myoblast differentiation in vitro and muscle regeneration in vivo

PLoS Genet. 2017 Feb 8;13(2):e1006600. doi: 10.1371/journal.pgen.1006600. eCollection 2017 Feb.

Abstract

The TEAD family of transcription factors (TEAD1-4) bind the MCAT element in the regulatory elements of both growth promoting and myogenic differentiation genes. Defining TEAD transcription factor function in myogenesis has proved elusive due to overlapping expression of family members and their functional redundancy. We show that silencing of either Tead1, Tead2 or Tead4 did not effect primary myoblast (PM) differentiation, but that their simultaneous knockdown strongly impaired differentiation. In contrast, Tead1 or Tead4 silencing impaired C2C12 differentiation showing their different contributions in PMs and C2C12 cells. Chromatin immunoprecipitation identified enhancers associated with myogenic genes bound by combinations of Tead4, Myod1 or Myog. Tead4 regulated distinct gene sets in C2C12 cells and PMs involving both activation of the myogenic program and repression of growth and signaling pathways. ChIP-seq from mature mouse muscle fibres in vivo identified a set of highly transcribed muscle cell-identity genes and sites bound by Tead1 and Tead4. Although inactivation of Tead4 in mature muscle fibres caused no obvious phenotype under normal conditions, notexin-induced muscle regeneration was delayed in Tead4 mutants suggesting an important role in myogenic differentiation in vivo. By combining knockdown in cell models in vitro with Tead4 inactivation in muscle in vivo, we provide the first comprehensive description of the specific and redundant roles of Tead factors in myogenic differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / genetics*
  • Cell Line
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Enhancer Elements, Genetic / genetics
  • Gene Expression Profiling / methods
  • Immunoblotting
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microscopy, Fluorescence
  • Muscle Development / genetics
  • Muscle Proteins / genetics*
  • Muscle Proteins / metabolism
  • Muscles / cytology
  • Muscles / metabolism*
  • Muscles / physiology
  • Mutation
  • Myoblasts / cytology
  • Myoblasts / metabolism*
  • Protein Binding
  • RNA Interference
  • Regeneration / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • DNA-Binding Proteins
  • Muscle Proteins
  • Tead1 protein, mouse
  • Tead4 protein, mouse
  • Transcription Factors

Grant support

This work was supported by grants from 1) The Association Française contre les Myopathies (AFM, http://www.afm-telethon.fr/association); 2) the Ligue Nationale contre le Cancer (https://www.ligue-cancer.net/); 3) the Agence National pour la Recherche (ANR, http://www.agence-nationale-recherche.fr/), Investissements d’Avenir program LABX-0030-INRT and 10-IDEX-0002-02; the Deutches Forschungs Gemeinschaft (http://www.dfg.de/) SFB TRR81, TPA2. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.