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Genome-wide analysis of FOXO1 binding features in early stage of chicken myoblast differentiation

ABSTRACT: Chromatin immunoprecipitation and high-throughput sequencing (ChIP-Seq) were performed for FOXO1 in early differentiated myoblasts. Based on FOXO1-occupied features, 234 coding genes were annotated including 39 targets overlapped with mouse ChIP-seq data. Function analysis demonstrated that these targets were enriched in myogenic proliferation, death and hypotrophy. Moreover, FOXO1 manipulated myoblast proliferation through promoting TGFBR2 and MYCN and repressing G0S2 at transcriptional level. On the other side, FOXO1 negatively regulated Bcl6 expression in myoblast, indicating that in the early stage of differentiation, myoblast apoptosis induced by miR-16 might be coordinated by impacts of FOXO1 on Bcl6.

ORGANISM(S): Gallus gallus

PROVIDER: GSE80533 | GEO | 2017/01/01

SECONDARY ACCESSION(S): PRJNA319207

REPOSITORIES: GEO

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Project description:Wnt/M-NM-2-catenin signaling is involved in various aspects of skeletal muscle development and regeneration. In addition, Wnt3a and M-NM-2-catenin are required for muscle-specific gene transcription in embryonic carcinoma cells and satellite-cell proliferation during adult skeletal muscle regeneration. Downstream targets of canonical Wnt signaling are cyclin D1 and c-myc. However, both target genes are suppressed during differentiation of mouse myoblast cells, C2C12. Underlying molecular mechanisms of M-NM-2-catenin signaling during myogenic differentiation remain unknown. Using C2C12 cells, we examined intracellular signaling and gene transcription during myoblast proliferation and differentiation. We confirmed that several Wnt signaling components, including Wnt9a, Sfrp2 and porcupine, were consistently upregulated in differentiating C2C12 cells. Troponin T-positive myotubes were decreased by Wnt3a overexpression, but not Wnt4. TOP/FOP reporter assays revealed that co-expression with Wnt4 reduced Wnt3a-induced luciferase activity, suggesting that Wnt4 signaling counteracted Wnt3a signaling in myoblasts. FH535, a small-molecule inhibitor of M-NM-2-catenin/Tcf complex formation, reduced basal M-NM-2-catenin in cytoplasm and decreased myoblast proliferation. K252a, a protein kinase inhibitor, increased membrane-bound M-NM-2-catenin and enhanced myoblast fusion. Treatments with K252a or Wnt4 resulted in increased cytoplasmic vesicles containing phosphorylated M-NM-2-catenin (Tyr654) during myogenic differentiation. These results suggest that various Wnt ligands control subcellular M-NM-2-catenin localization, which regulate myoblast proliferation and myotube formation. Wnt signaling via M-NM-2-catenin likely acts as a molecular switch that regulates the transition from cell proliferation to myogenic differentiation. Control cells (day 0) prior to differentiation induction with n=4; differentiated for two days with n=3; differentiated for four days with n=3.

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Genome-wide analysis of FOXO1 binding features in early stage of chicken myoblast differentiation

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