Cited by 304RNA Research and SplicingMicroRNA in disease regulationMuscle Physiology and Disorders

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MicroRNAs regulate the expression of the alternative splicing factor nPTB during muscle development

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Boutz, Paul L., Chawla, Geetanjali, Stoilov, Peter, Black, Douglas L.·2007·Genes & Development

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Abstract

Alternative pre-mRNA splicing determines many changes in gene expression during development. Two regulators known to control splicing patterns during neuron and muscle differentiation are the polypyrimidine tract-binding protein (PTB) and its neuronal homolog nPTB. These proteins repress certain exons in early myoblasts, but upon differentiation of mature myotubes PTB/nPTB expression is reduced, leading to increased inclusion of their target exons. We show here that the repression of nPTB expression during myoblast differentiation results from its targeting by the muscle-restricted microRNA miR-133. During differentiation of C2C12 myoblasts, nPTB protein but not mRNA expression is strongly reduced, concurrent with the up-regulation of miR-133 and the induction of splicing for several PTB-repressed exons. Introduction of synthetic miR-133 into undifferentiated C2C12 cells leads to a decrease in endogenous nPTB expression. Both the miR-133 and the coexpressed miR-1/206 microRNAs are extremely conserved across animal species, and PTB proteins are predicted targets for these miRNAs in Drosophila, mice, and humans. There are two potential miR-133-responsive elements (MRE) within the nPTB 3' untranslated region (UTR), and a luciferase reporter carrying this 3' UTR is repressed by miR-133 in an MRE-dependent manner. Transfection of locked nucleic acid (LNA) oligonucleotides designed to block the function of miR-133 and miR-1/206 increases expression of nPTB and decreases the inclusion of PTB dependent exons. These results indicate that miR-133 directly down-regulates a key splicing factor during muscle development and establishes a role for microRNAs in the control of a developmentally dynamic splicing program.

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Authors: Boutz, P. L., Chawla, G., Stoilov, P., & Black, D. L.
Affiliations: University of California, Los Angeles (US)
Howard Hughes Medical Institute (US)
ORCID: Boutz, Paul L.
Chawla, Geetanjali
Stoilov, Peter
Black, Douglas L.
Venue: Genes & Development
Publisher: Cold Spring Harbor Laboratory
Published: 2007-01-01
Volume: 21
Issue: 1
Pages: 71-84
ISSN: 0890-9369 (ISSN-L)
1549-5477
DOI: 10.1101/gad.1500707
PMC: PMC1759902
OpenAlex: W2166118364
Cited by: 304
References: 105
Citations by year: 2026: 4 · 2025: 1 · 2024: 3 · 2023: 9 · 2022: 9 · 2021: 5 · 2020: 10 · 2019: 10 · 2018: 14 · 2017: 10 · 2016: 14 · 2015: 13 · 2014: 16 · 2013: 24 · 2012: 18
Type: posted content
Language: en
Open access: bronze
doi.org
Locations: Genes & Development (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: Howard Hughes Medical Institute
National Institutes of Health (RO1 GM49662)
National Institute of General Medical Sciences (R01 GM 49662, R01GM049662)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
Keywords: microRNA, miR-133, polypyrimidine tract-binding protein, alternative splicing, myogenesis, 3' untranslated region, C2C12 myoblasts, LNA oligonucleotides
MeSH terms: Animals, Cell Differentiation, Cells, Cultured, Drosophila, Humans, Luciferases, Muscles, Neurons, Oligonucleotides, RNA Processing, Post-Transcriptional, Transfection, Blotting, Northern, Blotting, Western, Oligonucleotides, Antisense, Alternative Splicing, Gene Targeting, Gene Expression Regulation, Developmental, Reverse Transcriptase Polymerase Chain Reaction, Response Elements, 3' Untranslated Regions, Muscle Development, Myoblasts, RNA, Small Interfering, MicroRNAs, Polypyrimidine Tract-Binding Protein, Mice

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MicroRNAs regulate the expression of the alternative splicing factor nPTB during muscle development

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