Post-transcriptional regulation of MRTF-A by miRNAs during myogenic differentiation of myoblasts

Nucleic Acids Research
Ingo HolsteinGuido Posern

Abstract

The differentiation and regeneration of skeletal muscle from myoblasts to myotubes involves myogenic transcription factors, such as myocardin-related transcription factor A (MRTF-A) and serum response factor (SRF). In addition, post-transcriptional regulation by miRNAs is required during myogenesis. Here, we provide evidence for novel mechanisms regulating MRTF-A during myogenic differentiation. Endogenous MRTF-A protein abundance and activity decreased during C2C12 differentiation, which was attributable to miRNA-directed inhibition. Conversely, overexpression of MRTF-A impaired differentiation and myosin expression. Applying miRNA trapping by RNA affinity purification (miTRAP), we identified miRNAs which directly regulate MRTF-A via its 3'UTR, including miR-1a-3p, miR-206-3p, miR-24-3p and miR-486-5p. These miRNAs were upregulated during differentiation and specifically recruited to the 3'UTR of MRTF-A. Concomitantly, Ago2 recruitment to the MRTF-A 3'UTR was considerably increased, whereas Dicer1 depletion or 3'UTR deletion elevated MRTF-A and inhibited differentiation. MRTF-A protein expression was inhibited by ectopic miRNA expression in murine C2C12 and primary human myoblasts. 3'UTR reporter activity diminished upon diffe...Continue Reading

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Citations

Jul 3, 2021·International Journal of Molecular Sciences·Maria Zena MirandaAndrás Kapus
Aug 5, 2021·Journal of Cellular and Molecular Medicine·Ruhui SongYao Xu

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Datasets Mentioned

BETA
GSE136956

Methods Mentioned

BETA
GTPases
affinity purification
PCR
transfection
pulldown

Software Mentioned

bowtie
GraphPad prism
CASAVA
miRNome
edgeR
Image Studio
GraphPad
miTRAP

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