TEAD4 regulated genes in differentiated C2C12 myoblasts. Mus musculus

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 amongst the family members. Expression of the TEA/ATTS DNA binding domain that acts a dominant negative repressor of TEAD factors in C2C12 myoblasts inhibits their differentiation, while selective shRNA knockdown of TEAD4 results in abnormal differentiation characterised by the formation of shortened myotubes. RNA-seq identifies a novel set of TEAD4 target genes encoding muscle structural and regulatory proteins and those required for the unfolded protein response. In contrast, TEAD4 represses expression of the growth factor CTGF and Cyclin D1 to promote differentiation. Together these results show that TEAD factor activity is essential for normal C2C12 cell differentiation and define a novel and non-redundant role for TEAD4 in regulating expression of the unfolded protein response genes. Overall design: C2C12 cells were infected with lentiviral vectors expressing either a control shRNA or two independent shRNAs directed against TEAD4. The cell populations were selected with puromycin and then differentiated for 7 days to form myotubes.