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PMID: 17210790 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

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

Genes & development ·Vol. 21 ·No. 1 ·2007-01-01 ·Pages 71-84

Boutz PL, Chawla G, Stoilov P, Black DL

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.

MeSH Terms
3' Untranslated Regions/genetics Alternative Splicing Animals Blotting, Northern Blotting, Western Cell Differentiation Cells, Cultured Drosophila Gene Expression Regulation, Developmental Gene Targeting Humans Luciferases/metabolism Mice MicroRNAs/genetics,metabolism Muscle Development Muscles/metabolism Myoblasts/cytology Neurons/metabolism Oligonucleotides Oligonucleotides, Antisense/genetics Polypyrimidine Tract-Binding Protein/genetics RNA Processing, Post-Transcriptional RNA, Small Interfering/pharmacology Response Elements/genetics Reverse Transcriptase Polymerase Chain Reaction Transfection
Chemicals
3' Untranslated Regions MicroRNAs Oligonucleotides Oligonucleotides, Antisense RNA, Small Interfering locked nucleic acid Polypyrimidine Tract-Binding Protein Luciferases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Boutz Paul L
Department of Microbiology, Immunology, and Molecular Genetics, University of California at Los Angeles, Los Angeles, California 90095, USA.
Chawla Geetanjali
Stoilov Peter
Black Douglas L
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1500707
Published
2007-01-01
Pages
71-84
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC1759902
Subset
IM
Grants
NIGMS NIH HHS · R01 GM049662 · United States
NIGMS NIH HHS · R01 GM49662 · United States
Corrections
CommentIn
PMID: 17276332
External Links
PubMed source DOI: 10.1101/gad.1500707 PMC Full Text View journal details
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