title: Regulation of SIK1 abundance and stability is critical for myogenesis.
abstract: cAMP signaling can both promote and inhibit myogenic differentiation, but little is known about the mechanisms mediating promyogenic effects of cAMP. We previously demonstrated that the cAMP response element-binding protein (CREB) transcriptional target salt-inducible kinase 1 (SIK1) promotes MEF2 activity in myocytes via phosphorylation of class II histone deacetylase proteins (HDACs). However, it was unknown whether SIK1 couples cAMP signaling to the HDAC-MEF2 pathway during myogenesis and how this response could specifically occur in differentiating muscle cells. To address these questions, we explored SIK1 regulation and function in muscle precursor cells before and during myogenic differentiation. We found that in primary myogenic progenitor cells exposed to cAMP-inducing agents, Sik1 transcription is induced, but the protein is rapidly degraded by the proteasome. By contrast, sustained cAMP signaling extends the half-life of SIK1 in part by phosphorylation of Thr475, a previously uncharacterized site that we show can be phosphorylated by PKA in cell-free assays. We also identified a functional PEST domain near Thr475 that contributes to SIK1 degradation. During differentiation of primary myogenic progenitor cells, when PKA activity has been shown to increase, we observe elevated Sik1 transcripts as well as marked accumulation and stabilization of SIK1 protein. Depletion of Sik1 in primary muscle precursor cells profoundly impairs MEF2 protein accumulation and myogenic differentiation. Our findings support an emerging model in which SIK1 integrates cAMP signaling with the myogenic program to support appropriate timing of differentiation.
authors.affiliation: Graduate School of Biomedical Sciences, The University of Texas Medical School at Houston, Houston, TX 77030, USA.
authors.forename: Randi
authors.initials: R
authors.lastname: Stewart
authors.forename: Dmitry
authors.initials: D
authors.lastname: Akhmedov
authors.forename: Christopher
authors.initials: C
authors.lastname: Robb
authors.forename: Courtney
authors.initials: C
authors.lastname: Leiter
authors.forename: Rebecca
authors.initials: R
authors.lastname: Berdeaux
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D003720:Densitometry; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D057809:HEK293 Cells; D006207:Half-Life; D006801:Humans; D024510:Muscle Development / Q000502:physiology*; D032446:Myoblasts / Q000378:metabolism* / Q000502:physiology; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D017346:Protein Serine-Threonine Kinases; C509080:SIK1 protein, human; D046988:Proteasome Endopeptidase Complex