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p38 MAPK signaling regulates recruitment of Ash2L-containing methyltransferase complexes to specific genes during differentiation

Nature Structural & Molecular Biology volume 14, pages 1150–1156 (2007)Cite this article

Abstract

Cell-specific patterns of gene expression are established through the antagonistic functions of trithorax group (TrxG) and Polycomb group (PcG) proteins. Several muscle-specific genes have previously been shown to be epigenetically marked for repression by PcG proteins in muscle progenitor cells. Here we demonstrate that these developmentally regulated genes become epigenetically marked for gene expression (trimethylated on histone H3 Lys4, H3K4me3) during muscle differentiation through specific recruitment of Ash2L-containing methyltransferase complexes. Targeting of Ash2L to specific genes is mediated by the transcriptional regulator Mef2d. Furthermore, this interaction is modulated during differentiation through activation of the p38 MAPK signaling pathway via phosphorylation of Mef2d. Thus, we provide evidence that signaling pathways regulate the targeting of TrxG-mediated epigenetic modifications at specific promoters during cellular differentiation.

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Figure 1: Muscle-specific genes are trimethylated at H3K4 during myogenesis.
Figure 2: Mef2 interacts with the Ash2L methyltransferase complex.
Figure 3: Ash2L interacts preferentially with phosphorylated Mef2d and Mef2c.
Figure 4: Inhibition of p38 MAPK activity prevents recruitment of Ash2L to the Myog promoter.
Figure 5: Knockdown of Mef2d and Mef2c in C2C12 cells decreases Ash2L at the Myog and Ckm promoters.
Figure 6: Knockdown of Ash2L in C2C12 cells leads to reduced transcription of the Myog and Ckm genes.
Figure 7: Model for the integration of the p38 MAPK signaling pathway with Mef2-dependent recruitment of the Ash2L complex to muscle-specific promoters.

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References

  1. Ringrose, L. & Paro, R. Epigenetic regulation of cellular memory by the polycomb and trithorax group proteins. Annu. Rev. Genet. 38, 413–443 (2004).

    Article  CAS  Google Scholar 

  2. Brock, H.W. & Fisher, C.L. Maintenance of gene expression patterns. Dev. Dyn. 232, 633–655 (2005).

    Article  CAS  Google Scholar 

  3. Ruthenburg, A.J., Allis, C.D. & Wysocka, J. Methylation of lysine 4 on histone H3: intricacy of writing and reading a single epigenetic mark. Mol. Cell 25, 15–30 (2007).

    Article  CAS  Google Scholar 

  4. Bernstein, B.E. et al. Genomic maps and comparative analysis of histone modifications in human and mouse. Cell 120, 169–181 (2005).

    Article  CAS  Google Scholar 

  5. Martens, J.H. et al. The profile of repeat-associated histone lysine methylation states in the mouse epigenome. EMBO J. 24, 800–812 (2005).

    Article  CAS  Google Scholar 

  6. Papp, B. & Muller, J. Histone trimethylation and the maintenance of transcriptional ON and OFF states by trxG and PcG proteins. Genes Dev. 20, 2041–2054 (2006).

    Article  CAS  Google Scholar 

  7. Breen, T.R. & Harte, P.J. Molecular characterization of the trithorax gene, a positive regulator of homeotic gene expression in Drosophila. Mech. Dev. 35, 113–127 (1991).

    Article  CAS  Google Scholar 

  8. Klymenko, T. & Muller, J. The histone methyltransferases Trithorax and Ash1 prevent transcriptional silencing by Polycomb group proteins. EMBO Rep. 5, 373–377 (2004).

    Article  CAS  Google Scholar 

  9. Molkentin, J.D., Black, B.L., Martin, J.F. & Olson, E.N. Cooperative activation of muscle gene expression by MEf2 and myogenic bHLH proteins. Cell 83, 1125–1136 (1995).

    Article  CAS  Google Scholar 

  10. Bergstrom, D.A. et al. Promoter-specific regulation of MyoD binding and signal transduction cooperate to pattern gene expression. Mol. Cell 9, 587–600 (2002).

    Article  CAS  Google Scholar 

  11. Penn, B.H., Bergstrom, D.A., Dilworth, F.J., Bengal, E. & Tapscott, S.J.A. MyoD-generated feed forward circuit temporally patterns gene expression during skeletal muscle differentiation. Genes Dev. 18, 2348–2353 (2004).

    Article  CAS  Google Scholar 

  12. Wu, Z. et al. p38 and extracellular signal-regulated kinases regulate the myogenic program at multiple steps. Mol. Cell. Biol. 20, 3951–3964 (2000).

    Article  CAS  Google Scholar 

  13. Zetser, A., Gredinger, E. & Bengal, E. p38 mitogen-activated protein kinase pathway promotes skeletal muscle differentiation. Participation of the Mef2c transcription factor. J. Biol. Chem. 274, 5193–5200 (1999).

    Article  CAS  Google Scholar 

  14. Caretti, G., Di Padova, M., Micales, B., Lyons, G.E. & Sartorelli, V. The Polycomb Ezh2 methyltransferase regulates muscle gene expression and skeletal muscle differentiation. Genes Dev. 18, 2627–2638 (2004).

    Article  CAS  Google Scholar 

  15. Zhang, C.L., McKinsey, T.A. & Olson, E.N. Association of class II histone deacetylases with heterochromatin protein 1: potential role for histone methylation in control of muscle differentiation. Mol. Cell. Biol. 22, 7302–7312 (2002).

    Article  CAS  Google Scholar 

  16. Guenther, M.G., Levine, S.S., Boyer, L.A., Jaenisch, R. & Young, R.A. A chromatin landmark and transcription initiation at most promoters in human cells. Cell 130, 77–88 (2007).

    Article  CAS  Google Scholar 

  17. Yaffe, D. & Saxel, O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature 270, 725–727 (1977).

    Article  CAS  Google Scholar 

  18. Milne, T.A. et al. MLL associates specifically with a subset of transcriptionally active target genes. Proc. Natl. Acad. Sci. USA 102, 14765–14770 (2005).

    Article  CAS  Google Scholar 

  19. Steward, M.M. et al. Molecular regulation of H3K4 trimethylation by ASH2L, a shared subunit of MLL complexes. Nat. Struct. Mol. Biol. 13, 852–854 (2006).

    Article  CAS  Google Scholar 

  20. Dou, Y. et al. Regulation of MLL1 H3K4 methyltransferase activity by its core components. Nat. Struct. Mol. Biol. 13, 713–719 (2006).

    Article  CAS  Google Scholar 

  21. Wang, J. et al. ASH2L: alternative splicing and downregulation during induced megakaryocytic differentiation of multipotential leukemia cell lines. J. Mol. Med. 79, 399–405 (2001).

    Article  CAS  Google Scholar 

  22. Cheng, T.C., Wallace, M.C., Merlie, J.P. & Olson, E.N. Separable regulatory elements governing myogenin transcription in mouse embryogenesis. Science 261, 215–218 (1993).

    Article  CAS  Google Scholar 

  23. Yee, S.P. & Rigby, P.W. The regulation of myogenin gene expression during the embryonic development of the mouse. Genes Dev. 7, 1277–1289 (1993).

    Article  CAS  Google Scholar 

  24. Edmondson, D.G., Cheng, T.C., Cserjesi, P., Chakraborty, T. & Olson, E.N. Analysis of the myogenin promoter reveals an indirect pathway for positive autoregulation mediated by the muscle-specific enhancer factor MEF-2. Mol. Cell. Biol. 12, 3665–3677 (1992).

    Article  CAS  Google Scholar 

  25. Malik, S., Huang, C.F. & Schmidt, J. The role of the CANNTG promoter element (E box) and the myocyte-enhancer-binding-factor-2 (MEF-2) site in the transcriptional regulation of the chick myogenin gene. Eur. J. Biochem. 230, 88–96 (1995).

    Article  CAS  Google Scholar 

  26. Berkes, C.A. et al. Pbx marks genes for activation by MyoD indicating a role for a homeodomain protein in establishing myogenic potential. Mol. Cell 14, 465–477 (2004).

    Article  CAS  Google Scholar 

  27. Cho, Y.W. et al. PTIP associates with MLL3- and MLL4-containing histone H3 lysine 4 methyltransferase complex. J. Biol. Chem. 282, 20395–20406 (2007).

    Article  CAS  Google Scholar 

  28. Zhao, M. et al. Regulation of the MEf2 family of transcription factors by p38. Mol. Cell. Biol. 19, 21–30 (1999).

    Article  CAS  Google Scholar 

  29. Khokhlatchev, A. et al. Reconstitution of mitogen-activated protein kinase phosphorylation cascades in bacteria. Efficient synthesis of active protein kinases. J. Biol. Chem. 272, 11057–11062 (1997).

    Article  CAS  Google Scholar 

  30. Zhu, B., Ramachandran, B. & Gulick, T. Alternative pre-mRNA splicing governs expression of a conserved acidic transactivation domain in myocyte enhancer factor 2 factors of striated muscle and brain. J. Biol. Chem. 280, 28749–28760 (2005).

    Article  CAS  Google Scholar 

  31. Simone, C. et al. p38 pathway targets SWI-SNF chromatin-remodeling complex to muscle-specific loci. Nat. Genet. 36, 738–743 (2004).

    Article  CAS  Google Scholar 

  32. Ernst, P., Wang, J., Huang, M., Goodman, R.H. & Korsmeyer, S.J. MLL and CREB bind cooperatively to the nuclear coactivator CREB-binding protein. Mol. Cell. Biol. 21, 2249–2258 (2001).

    Article  CAS  Google Scholar 

  33. Voo, K.S., Carlone, D.L., Jacobsen, B.M., Flodin, A. & Skalnik, D.G. Cloning of a mammalian transcriptional activator that binds unmethylated CpG motifs and shares a CXXC domain with DNA methyltransferase, human trithorax, and methyl-CpG binding domain protein 1. Mol. Cell. Biol. 20, 2108–2121 (2000).

    Article  CAS  Google Scholar 

  34. Wysocka, J. et al. WDR5 associates with histone H3 methylated at K4 and is essential for H3 K4 methylation and vertebrate development. Cell 121, 859–872 (2005).

    Article  CAS  Google Scholar 

  35. Lluis, F., Perdiguero, E. & Nebreda, A.R. Munoz-Canoves, P. Regulation of skeletal muscle gene expression by p38 MAP kinases. Trends Cell Biol. 16, 36–44 (2006).

    Article  CAS  Google Scholar 

  36. Takaesu, G. et al. Activation of p38α/β MAPK in myogenesis via binding of the scaffold protein JLP to the cell surface protein Cdo. J. Cell Biol. 175, 383–388 (2006).

    Article  CAS  Google Scholar 

  37. Ohkawa, Y., Marfella, C.G. & Imbalzano, A.N. Skeletal muscle specification by myogenin and Mef2D via the SWI/SNF ATPase Brg1. EMBO J. 25, 490–501 (2006).

    Article  CAS  Google Scholar 

  38. de la Serna, I.L. et al. MyoD targets chromatin remodeling complexes to the myogenin locus prior to forming a stable DNA-bound complex. Mol. Cell. Biol. 25, 3997–4009 (2005).

    Article  CAS  Google Scholar 

  39. Brand, M. et al. Dynamic changes in transcription factor complexes during erythroid differentiation revealed by quantitative proteomics. Nat. Struct. Mol. Biol. 11, 73–80 (2004).

    Article  CAS  Google Scholar 

  40. Dilworth, F.J., Ramain-Fromental, C., Yamamoto, K. & Chambon, P. ATP-driven chromatin remodeling activity and histone acetyltransferases act sequencially during transactivation by RAR/RXR in vitro. Mol. Cell 6, 1049–1058 (2000).

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by grants from the Canadian Institutes of Health Research, the Muscular Dystrophy Association and the Stem Cell Network (to F.J.D.), from the US National Institutes of Health (to S.J.T.) and from the Terry Fox Foundation of the National Cancer Institute of Canada and the Human Frontiers Science Program (to M.B.). F.J.D. holds a Canadian Research Chair in Epigenetic Regulation of Transcription.

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Authors and Affiliations

  1. Sprott Center for Stem Cell Research, Ottawa Health Research Institute, Ottawa, K1H 8L6, Ontario, Canada

    Shravanti Rampalli, LiFang Li, Esther Mak, Marjorie Brand & F Jeffrey Dilworth

  2. National Institute of Diabetes and Digestive and Kidney Diseases, US National Institutes of Health, Bethesda, 20892, Maryland, USA

    Kai Ge

  3. Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, K1H 8L6, Ontario, Canada

    Marjorie Brand & F Jeffrey Dilworth

  4. Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, 98109, Washington, USA

    Stephen J Tapscott

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Contributions

S.R. and F.J.D. conceived and designed the experiments. S.R. was responsible for all ChIP, immunoprecipitation, RT-PCR and interaction studies. F.J.D. and M.B. generated the polyclonal antibody to full-length Ash2L. L.L. generated the polyclonal antibodies to full-length DPY and MyoD. E.M. generated the Mef2D mutant. S.R. and E.M. performed the transfection studies with the Mef2 mutants. K.G. provided antibodies to MLL3 and MLL4. S.R., M.B., S.J.T. and F.J.D. provided scientific direction of the project. F.J.D. wrote the paper. S.R., M.B. and F.J.D. discussed and commented on the manuscript.

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Correspondence to F Jeffrey Dilworth.

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Rampalli, S., Li, L., Mak, E. et al. p38 MAPK signaling regulates recruitment of Ash2L-containing methyltransferase complexes to specific genes during differentiation. Nat Struct Mol Biol 14, 1150–1156 (2007). https://doi.org/10.1038/nsmb1316

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