TBP/TFIID-dependent activation of MyoD target genes in skeletal muscle cells.

ELife
Barbora MalecovaPier Lorenzo Puri

Abstract

Change in the identity of the components of the transcription pre-initiation complex is proposed to control cell type-specific gene expression. Replacement of the canonical TFIID-TBP complex with TRF3/TBP2 was reported to be required for activation of muscle-gene expression. The lack of a developmental phenotype in TBP2 null mice prompted further analysis to determine whether TBP2 deficiency can compromise adult myogenesis. We show here that TBP2 null mice have an intact regeneration potential upon injury and that TBP2 is not expressed in established C2C12 muscle cell or in primary mouse MuSCs. While TFIID subunits and TBP are downregulated during myoblast differentiation, reduced amounts of these proteins form a complex that is detectable on promoters of muscle genes and is essential for their expression. This evidence demonstrates that TBP2 does not replace TBP during muscle differentiation, as previously proposed, with limiting amounts of TFIID-TBP being required to promote muscle-specific gene expression.

References

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Methods Mentioned

BETA
Fluorescence Assisted Cell Sorting
RNA-seq
Immunoprecipitation
co-immunoprecipitation
size fractionation assay
fluorescence-activated cell sorting
Flow Cytometry
FACS
transfection
PCR

Software Mentioned

FACSDiva
TOPHAT
ImageJ
Zen
UCSC genome browser

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