Abstract
The TEAD (1–4) transcription factors comprise the conserved TEA/ATTS DNA-binding domain recognising the MCAT element in the promoters of muscle-specific genes. Despite extensive genetic analysis, the function of TEAD factors in muscle differentiation has proved elusive due to redundancy among the family members. Expression of the TEA/ATTS DNA-binding domain that acts as a dominant negative repressor of TEAD factors in C2C12 myoblasts inhibits their differentiation, whereas selective shRNA knockdown of TEAD4 results in abnormal differentiation characterised by the formation of shortened myotubes. Chromatin immunoprecipitation coupled to array hybridisation shows that TEAD4 occupies 867 promoters including those of myogenic miRNAs. We show that TEAD factors directly induce Myogenin, CDKN1A and Caveolin 3 expression to promote myoblast differentiation. RNA-seq identifies a set of genes whose expression is strongly reduced upon TEAD4 knockdown among which are structural and regulatory proteins and those required for the unfolded protein response. In contrast, TEAD4 represses expression of the growth factor CTGF (connective tissue growth factor) to promote differentiation. Together these results show that TEAD factor activity is essential for normal C2C12 cell differentiation and suggest a role for TEAD4 in regulating expression of the unfolded protein response genes.
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Abbreviations
- TEA:
-
Yeast (TEC-1), Aspergillus nidulans (AbaA) and Drosophilla (scalloped)
- ATTS:
-
Aspergillus nidulans (AbaA), Yeast (TEC-1), human TEF1, and Drosophilla (scalloped)
- DBD:
-
DNA-binding domain
- SV40:
-
simian virus 40
- CTGF:
-
connective tissue growth factor
- β-MHC, myosin, heavy polypeptide 7, cardiac muscle, beta; YAP1:
-
Yes-associated protein 1
- WWTR1:
-
WW domain containing transcription regulator 1
- ChIP-chip:
-
chromatin immunoprecipitation-array hybridization
- Flag:
-
FLAG octapeptide tag
- HA:
-
Hemagglutinin tag
- TSS:
-
transcription start site
- UPR:
-
unfolded protein response
- Cdkn1a:
-
cyclin-dependent kinase inhibitor 1A (P21)
- TRP53:
-
transformation-related protein 53
- SP1:
-
trans-acting transcription factor 1
- RT-qPCR:
-
reverse transcription–quantitative polymerase chain reaction
- VGL:
-
vestigial
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Acknowledgements
We thank K Hnia and J Laporte for the gift of antibodies and advice. This work was supported by grants from the CNRS, the INSERM, the Université de Strasbourg, the Association pour la Recherche contre le Cancer, the Ligue Nationale contre le Cancer, the INCA and the AFM. ID is an ‘équipe labellisée’ of the Ligue Nationale contre le Cancer. AB was supported by a followship from the AFM. The mRNA-seq data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE27845 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE27845). The ChIP-chip data are are accessible through GEO Series accession number GSE29208.
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Benhaddou, A., Keime, C., Ye, T. et al. Transcription factor TEAD4 regulates expression of Myogenin and the unfolded protein response genes during C2C12 cell differentiation. Cell Death Differ 19, 220–231 (2012). https://doi.org/10.1038/cdd.2011.87
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DOI: https://doi.org/10.1038/cdd.2011.87
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