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Alternatively spliced exon regulates context-dependent MEF2D higher-order assembly during myogenesis.

Mónika Gönczi1,2, João M C Teixeira3, Susana Barrera-Vilarmau3

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Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Muscle cell differentiation involves complex transcriptional regulation.
  • Myocyte-specific Enhancer Factor 2 (Mef2D) plays a crucial role in this process.
  • Alternative splicing of Mef2D generates a β-domain that influences its function.

Purpose of the Study:

  • To investigate the role of the Mef2D β-domain in higher-order protein assembly.
  • To explore the relationship between Mef2D assembly states and transcriptional activity.
  • To understand how the β-domain's properties affect Mef2D's cellular localization and function during myogenesis.

Main Methods:

  • Sequence analysis using the FuzDrop method.
  • Live-cell imaging of Mef2D condensates and aggregates in C2C12 cells.
  • Analysis of MyoD and desmin expression.
  • Biophysical characterization of β-domain variants.
  • Nuclear Magnetic Resonance (NMR) spectroscopy.
  • Molecular dynamics (MD) simulations.

Main Results:

  • The Mef2D β-domain acts as an interaction element for Mef2D higher-order assembly.
  • Mef2D forms mobile nuclear condensates and solid-like cytosolic aggregates.
  • Cytosolic Mef2D aggregates correlate with increased transcriptional activity and enhanced myotube development.
  • β-domain variants influence the formation of liquid-like and solid-like Mef2D assemblies.
  • NMR and MD simulations confirm the β-domain's ability to adopt ordered and disordered conformations.

Conclusions:

  • The Mef2D β-domain dynamically regulates Mef2D higher-order assembly.
  • Mef2D assembly states are fine-tuned to the cellular context, impacting transcriptional regulation.
  • This regulation provides a platform for myogenic factors and the transcriptional machinery during muscle development.