1. Phosphorylation of TET2 by AMPK is indispensable in myogenic differentiation.
- Author
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Zhang T, Guan X, Choi UL, Dong Q, Lam MMT, Zeng J, Xiong J, Wang X, Poon TCW, Zhang H, Zhang X, Wang H, Xie R, Zhu B, and Li G
- Subjects
- 14-3-3 Proteins metabolism, AMP-Activated Protein Kinases genetics, Animals, Cell Differentiation genetics, DNA Methylation, DNA-Binding Proteins genetics, Dioxygenases, Gene Knockout Techniques, Genome-Wide Association Study, HEK293 Cells, Humans, Mice, Muscle Fibers, Skeletal cytology, Muscle Fibers, Skeletal metabolism, Myoblasts cytology, Myoblasts metabolism, PAX7 Transcription Factor biosynthesis, PAX7 Transcription Factor genetics, PAX7 Transcription Factor metabolism, Phosphorylation, Protein Binding, Proto-Oncogene Proteins genetics, AMP-Activated Protein Kinases metabolism, DNA-Binding Proteins metabolism, Muscle Development physiology, Muscles cytology, Muscles metabolism, Proto-Oncogene Proteins metabolism
- Abstract
Background: TET-mediated oxidation of 5-mC participates in both passive and active DNA demethylation, which exerts a significant influence on diverse biological processes. Mass spectrometry has identified multiple phosphorylation sites of TET2. However, the functions of these phosphosites and their corresponding kinases are mostly unknown., Results: Here, we showed that AMP-activated protein kinase (AMPK) phosphorylates murine TET2 at the serine residue 97 (S97), and the phosphorylation enhances TET2 stability through promoting its binding to 14-3-3β. AMPK ablation resulted in decreased global 5-hmC levels at the myotube stages, severe differentiation defects of C2C12 cells and significantly, total loss of expression of Pax7. Genome-wide analyses revealed increased DNA methylation at genic and enhancer regions of AMPK-null myoblasts and myotubes. Using CRISPR/Cas9 technology, we showed that a novel enhancer, which is hypermethylated in AMPK-null cells, regulates Pax7 expression. The phospho-mimicking mutant, TET2-S97E, could partly rescue the differentiation defect in AMPK-ablated C2C12 cells., Conclusions: Together, our data demonstrated that AMPK is a critical regulator of myogenesis, partly through phosphorylating TET2.
- Published
- 2019
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