Ythdf2 facilitates precursor miR-378/miR-378-5p maturation to support myogenic differentiation

Area of Science:

• Molecular Biology
• Epigenetics
• Cell Biology

Background:

• Ythdf2 protein mediates mRNA degradation and influences skeletal muscle differentiation.
• Ythdf2 interacts with m6A-modified precursor miRNAs, affecting their maturation.
• The role of Ythdf2 in myogenesis via miRNA maturation remains unclear.

Purpose of the Study:

• To investigate whether Ythdf2 regulates myogenesis by controlling miRNA maturation.
• To identify specific miRNAs targeted by Ythdf2 during myogenic differentiation.
• To elucidate the molecular mechanism linking Ythdf2, miRNA maturation, and myogenesis.

Main Methods:

• Ythdf2 knockdown experiments in myoblasts.
• miRNA and mRNA sequencing for integrated analysis.
• Co-immunoprecipitation assays to study protein interactions.
• Western blotting and qRT-PCR to assess protein and miRNA expression.

Main Results:

• Ythdf2 knockdown suppressed myotube formation and altered miRNA expression.
• miR-378 and miR-378-5p were identified as key Ythdf2 targets in myogenesis.
• Ythdf2 facilitates pre-miR-378/miR-378-5p maturation by interacting with DICER1 and TARBP2.
• Downregulation of miR-378/miR-378-5p inhibited myogenesis; their forced expression rescued Ythdf2 knockdown effects via the mTOR pathway.

Conclusions:

• Ythdf2 regulates myogenic differentiation by mediating pre-miR-378/miR-378-5p maturation in an m6A-dependent manner.
• This mechanism involves Ythdf2 interaction with the pre-miRNA processing complex.
• The findings provide novel insights into m6A modification's role in myogenesis regulation.