Abstract
Transforming growth factor-beta (TGF-beta) and insulin-like growth factors (IGFs) play critical roles in the control of myogenesis. Insulin-like growth factor-binding protein-5 (IGFBP-5), by regulating the bioavailability of IGFs, is involved in controlling IGF-dependent differentiation. We investigated the effects of TGF-beta on the IGFBP-5 production induced by IGFs in mouse myoblasts. TGF-beta leads to a decrease in IGFBP-5 synthesis at both transcript and protein levels, and blocked muscle differentiation. The Smad proteins and the c-Jun N-terminal kinase (JNK) have been shown to be involved in TGF-beta signaling pathways. We provide evidence that the JNK pathway, rather than Smad proteins, is involved in the response of muscle cells to TGF-beta. This factor failed to stimulate the GAL4-Smad 2/3 transcriptional activities of the constructs used to transfect myoblasts. Moreover, stable expression of the antagonistic Smad7 did not abolish the inhibitory effect of TGF-beta on IGFBP-5 production whereas expression of a dominant-negative version of MKK4, an upstream activator of JNK, did. We also showed, using a specific inhibitor, that the p38 mitogen-activated protein kinase (p38 MAPK) was not involved in the inhibition of IGFBP-5 production. Thus, TGF-beta-mediated IGFBP-5 inhibition is independent of Smads and requires activation of the JNK signaling pathway.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Apoptosis
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Blotting, Northern
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Cell Differentiation
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Cells, Cultured
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DNA-Binding Proteins / metabolism
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Dose-Response Relationship, Drug
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Enzyme Activation
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Genes, Dominant
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Insulin / metabolism
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Insulin-Like Growth Factor Binding Protein 5 / antagonists & inhibitors*
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Insulin-Like Growth Factor Binding Protein 5 / biosynthesis
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Insulin-Like Growth Factor Binding Protein 5 / metabolism*
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JNK Mitogen-Activated Protein Kinases
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Luciferases / metabolism
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MAP Kinase Kinase 4*
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Mice
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Mitogen-Activated Protein Kinase Kinases / genetics
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Mitogen-Activated Protein Kinase Kinases / metabolism
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Mitogen-Activated Protein Kinases / metabolism*
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Muscle, Skeletal / metabolism*
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Mutation
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Plasmids / metabolism
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Protein Binding
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RNA, Messenger / metabolism
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Signal Transduction
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Smad2 Protein
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Smad3 Protein
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Smad7 Protein
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Trans-Activators / metabolism
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Transcription, Genetic
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Transcriptional Activation
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Transfection
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Transforming Growth Factor beta / metabolism*
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Troponin T / metabolism
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p38 Mitogen-Activated Protein Kinases
Substances
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DNA-Binding Proteins
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Insulin
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Insulin-Like Growth Factor Binding Protein 5
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RNA, Messenger
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Smad2 Protein
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Smad2 protein, mouse
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Smad3 Protein
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Smad3 protein, mouse
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Smad7 Protein
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Smad7 protein, mouse
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Trans-Activators
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Transforming Growth Factor beta
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Troponin T
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Luciferases
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JNK Mitogen-Activated Protein Kinases
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Mitogen-Activated Protein Kinases
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p38 Mitogen-Activated Protein Kinases
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MAP Kinase Kinase 4
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Map2k4 protein, mouse
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Mitogen-Activated Protein Kinase Kinases