Time-dependent Pax3-mediated chromatin remodeling and cooperation with Six4 and Tead2 specify the skeletal myogenic lineage in developing mesoderm

PLoS Biol. 2019 Feb 26;17(2):e3000153. doi: 10.1371/journal.pbio.3000153. eCollection 2019 Feb.


The transcriptional mechanisms driving lineage specification during development are still largely unknown, as the interplay of multiple transcription factors makes it difficult to dissect these molecular events. Using a cell-based differentiation platform to probe transcription function, we investigated the role of the key paraxial mesoderm and skeletal myogenic commitment factors-mesogenin 1 (Msgn1), T-box 6 (Tbx6), forkhead box C1 (Foxc1), paired box 3 (Pax3), Paraxis, mesenchyme homeobox 1 (Meox1), sine oculis-related homeobox 1 (Six1), and myogenic factor 5 (Myf5)-in paraxial mesoderm and skeletal myogenesis. From this study, we define a genetic hierarchy, with Pax3 emerging as the gatekeeper between the presomitic mesoderm and the myogenic lineage. By assaying chromatin accessibility, genomic binding and transcription profiling in mesodermal cells from mouse and human Pax3-induced embryonic stem cells and Pax3-null embryonic day (E)9.5 mouse embryos, we identified conserved Pax3 functions in the activation of the skeletal myogenic lineage through modulation of Hedgehog, Notch, and bone morphogenetic protein (BMP) signaling pathways. In addition, we demonstrate that Pax3 molecular function involves chromatin remodeling of its bound elements through an increase in chromatin accessibility and cooperation with sine oculis-related homeobox 4 (Six4) and TEA domain family member 2 (Tead2) factors. To our knowledge, these data provide the first integrated analysis of Pax3 function, demonstrating its ability to remodel chromatin in mesodermal cells from developing embryos and proving a mechanistic footing for the transcriptional hierarchy driving myogenesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Differentiation
  • Cell Line
  • Chromatin Assembly and Disassembly*
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Embryo, Mammalian
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Forkhead Transcription Factors / genetics
  • Forkhead Transcription Factors / metabolism
  • Gene Expression Regulation, Developmental
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism
  • Humans
  • Mesoderm / cytology
  • Mesoderm / growth & development
  • Mesoderm / metabolism*
  • Mice
  • Mice, Transgenic
  • Muscle Cells / cytology
  • Muscle Cells / metabolism*
  • Muscle Development / genetics*
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / growth & development
  • Muscle, Skeletal / metabolism
  • Myogenic Regulatory Factor 5 / genetics
  • Myogenic Regulatory Factor 5 / metabolism
  • PAX3 Transcription Factor / genetics*
  • PAX3 Transcription Factor / metabolism
  • Signal Transduction
  • T-Box Domain Proteins
  • TEA Domain Transcription Factors
  • Trans-Activators / genetics*
  • Trans-Activators / metabolism
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism


  • Basic Helix-Loop-Helix Transcription Factors
  • DNA-Binding Proteins
  • Forkhead Transcription Factors
  • Foxc1 protein, mouse
  • Homeodomain Proteins
  • Meox1 protein, mouse
  • Msgn1 protein, mouse
  • Myf5 protein, mouse
  • Myogenic Regulatory Factor 5
  • PAX3 Transcription Factor
  • Six1 protein, mouse
  • Six4 protein, mouse
  • T-Box Domain Proteins
  • TEA Domain Transcription Factors
  • Tbx6 protein, mouse
  • Tead2 protein, mouse
  • Trans-Activators
  • Transcription Factors
  • Pax3 protein, mouse