Program in Molecular Medicine
Animals; Cell Differentiation; Cell Line; *Gene Expression Regulation, Developmental; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Muscle Development; Muscle Fibers, Skeletal; Myoblasts; Myogenic Regulatory Factor 5; Protein-Serine-Threonine Kinases; RNA Interference; RNA, Small Interfering; Up-Regulation
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Myoblast differentiation into mature myotubes is a critical step in the development and repair of human skeletal muscle. Here we show that small interfering RNA (siRNA)-based silencing of the Ste20-like mitogen-activated protein 4 kinase 4 (Map4k4) in C2C12 myoblasts markedly enhances expression of myogenic differentiation genes, myoblast fusion, and myotube diameter. In contrast, adenovirus-mediated expression of native Map4k4 in C2C12 cells attenuates each of these processes, indicating that Map4k4 is a negative regulator of myogenic differentiation and hypertrophy. Expression of a Map4k4 kinase-inactive mutant enhances myotube formation, suggesting that the kinase activity of Map4k4 is essential for its inhibition of muscle differentiation. Map4k4 regulation of myogenesis is unlikely to be mediated by classic mitogen-activated protein kinase (MAPK) signaling pathways, because no significant difference in phosphorylation of extracellular signal-regulated kinase (ERK), p38, or c-Jun N-terminal kinase (JNK) is observed in Map4k4-silenced cells. Furthermore, silencing of these other MAPKs does not result in a hypertrophic myotube phenotype like that seen with Map4k4 depletion. Uniquely, Map4k4 silencing upregulates the expression of the myogenic regulatory factor Myf5, whose depletion inhibits myogenesis. Furthermore, Myf5 is required for enhancement of myotube formation in Map4k4-silenced cells, while Myf5 overexpression rescues Map4k4-mediated inhibition of myogenic differentiation. These results demonstrate that Map4k4 is a novel suppressor of skeletal muscle differentiation, acting through a Myf5-dependent mechanism.
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DOI of Published Version
Wang M, Amano SU, Flach RJ, Chawla A, Aouadi M, Czech MP. Identification of Map4k4 as a novel suppressor of skeletal muscle differentiation. Mol Cell Biol. 2013 Feb;33(4):678-87. doi: 10.1128/MCB.00618-12. Link to article on publisher's site
Molecular and cellular biology
Wang M, Amano SU, Roth Flach RJ, Chawla A, Aouadi M, Czech MP. (2013). Identification of Map4k4 as a novel suppressor of skeletal muscle differentiation. University of Massachusetts Medical School Faculty Publications. https://doi.org/10.1128/MCB.00618-12. Retrieved from https://escholarship.umassmed.edu/faculty_pubs/199
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