Opposing calcium-dependent signalling pathways control skeletal muscle differentiation by regulating a chromatin remodelling enzyme
Authors
Nasipak, Brian T.Padilla-Benavides, Teresita
Green, Karin M.
Leszyk, John D.
Mao, Wenjie
Konda, Silvana
Sif, Said
Shaffer, Scott A.
Ohkawa, Yasuyuki
Imbalzano, Anthony N.
Student Authors
Wenjie MaoAcademic Program
NeuroscienceUMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyProteomics and Mass Spectrometry Facility
Department of Cell and Developmental Biology
Document Type
Journal ArticlePublication Date
2015-06-17Keywords
Biological sciencesCell biology
Developmental biology
Cell Biology
Computational Biology
Developmental Biology
Metadata
Show full item recordAbstract
Calcium signalling is important for differentiation-dependent gene expression, but is also involved in other cellular functions. Therefore, mechanisms must exist to distinguish calcium signalling relevant to differentiation. Calcineurin is a calcium-regulated phosphatase that is required for myogenic gene expression and skeletal muscle differentiation. Here, we demonstrate that inhibition of calcineurin blocks chromatin remodelling and that the Brg1 ATPase of the SWI/SNF chromatin remodelling enzyme, which is required for the activation of myogenic gene expression, is a calcineurin substrate. Furthermore, we identify the calcium-regulated classical protein kinase C beta (PKCbeta) as a repressor of myogenesis and as the enzyme that opposes calcineurin function. Replacement of endogenous Brg1 with a phosphomimetic mutant in primary myoblasts inhibits myogenesis, whereas replacement with a non-phosphorylatable mutant allows myogenesis despite inhibition of calcineurin signalling, demonstrating the functionality of calcineurin/PKC-modified residues. Thus, the Brg1 chromatin remodelling enzyme integrates two antagonistic calcium-dependent signalling pathways that control myogenic differentiation.Source
Nat Commun. 2015 Jun 17;6:7441. doi: 10.1038/ncomms8441. Link to article on publisher's siteDOI
10.1038/ncomms8441Permanent Link to this Item
http://hdl.handle.net/20.500.14038/26483PubMed ID
26081415Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1038/ncomms8441
Scopus Count
Collections
Related items
Showing items related by title, author, creator and subject.
-
Mapping and analysis of Caenorhabditis elegans transcription factor sequence specificitiesNarasimhan, Kamesh; Lambert, Samuel A.; Yang, Ally; Riddell, Jeremy; Mnaimneh, Sanie; Zheng, Hong; Albu, Mihai; Najafabadi, Hamed S.; Reece-Hoyes, John S.; Fuxman Bass, Juan; et al. (2015-04-23)Caenorhabditis elegans is a powerful model for studying gene regulation, as it has a compact genome and a wealth of genomic tools. However, identification of regulatory elements has been limited, as DNA-binding motifs are known for only 71 of the estimated 763 sequence-specific transcription factors (TFs). To address this problem, we performed protein binding microarray experiments on representatives of canonical TF families in C. elegans, obtaining motifs for 129 TFs. Additionally, we predict motifs for many TFs that have DNA-binding domains similar to those already characterized, increasing coverage of binding specificities to 292 C. elegans TFs (~40%). These data highlight the diversification of binding motifs for the nuclear hormone receptor and C2H2 zinc finger families, and reveal unexpected diversity of motifs for T-box and DM families. Motif enrichment in promoters of functionally related genes is consistent with known biology, and also identifies putative regulatory roles for unstudied TFs.
-
Combined experimental and computational analysis of DNA damage signaling reveals context-dependent roles for Erk in apoptosis and G1/S arrest after genotoxic stressTentner, Andrea R.; Lee, Michael J; Ostheimer, Gerry J.; Samson, Leona D.; Lauffenburger, Douglas A.; Yaffe, Michael B. (2012-01-31)Following DNA damage, cells display complex multi-pathway signaling dynamics that connect cell-cycle arrest and DNA repair in G1, S, or G2/M phase with phenotypic fate decisions made between survival, cell-cycle re-entry and proliferation, permanent cell-cycle arrest, or cell death. How these phenotypic fate decisions are determined remains poorly understood, but must derive from integrating genotoxic stress signals together with inputs from the local microenvironment. To investigate this in a systematic manner, we undertook a quantitative time-resolved cell signaling and phenotypic response study in U2OS cells receiving doxorubicin-induced DNA damage in the presence or absence of TNFalpha co-treatment; we measured key nodes in a broad set of DNA damage signal transduction pathways along with apoptotic death and cell-cycle regulatory responses. Two relational modeling approaches were then used to identify network-level relationships between signals and cell phenotypic events: a partial least squares regression approach and a complementary new technique which we term 'time-interval stepwise regression.' Taken together, the results from these analysis methods revealed complex, cytokine-modulated inter-relationships among multiple signaling pathways following DNA damage, and identified an unexpected context-dependent role for Erk in both G1/S arrest and apoptotic cell death following treatment with this commonly used clinical chemotherapeutic drug.
-
Determination of ubiquitin fitness landscapes under different chemical stresses in a classroom settingMavor, David; Roscoe, Benjamin P.; Bolon, Daniel N.; Fraser, James S. (2016-04-25)Ubiquitin is essential for eukaryotic life and varies in only 3 amino acid positions between yeast and humans. However, recent deep sequencing studies indicate that ubiquitin is highly tolerant to single mutations. We hypothesized that this tolerance would be reduced by chemically induced physiologic perturbations. To test this hypothesis, a class of first year UCSF graduate students employed deep mutational scanning to determine the fitness landscape of all possible single residue mutations in the presence of five different small molecule perturbations. These perturbations uncover 'shared sensitized positions' localized to areas around the hydrophobic patch and the C-terminus. In addition, we identified perturbation specific effects such as a sensitization of His68 in HU and a tolerance to mutation at Lys63 in DTT. Our data show how chemical stresses can reduce buffering effects in the ubiquitin proteasome system. Finally, this study demonstrates the potential of lab-based interdisciplinary graduate curriculum.