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Pax7 activates myogenic genes by recruitment of a histone methyltransferase complex

Nature Cell Biology volume 10, pages 77–84 (2008)Cite this article

Abstract

Satellite cells purified from adult skeletal muscle can participate extensively in muscle regeneration and can also re-populate the satellite cell pool, suggesting that they have direct therapeutic potential for treating degenerative muscle diseases1,2. The paired-box transcription factor Pax7 is required for satellite cells to generate committed myogenic progenitors3. In this study we undertook a multi-level approach to define the role of Pax7 in satellite cell function. Using comparative microarray analysis, we identified several novel and strongly regulated targets; in particular, we identified Myf5 as a gene whose expression was regulated by Pax7. Using siRNA, fluorescence-activated cell sorting (FACS) and chromatin immunoprecipitation (ChIP) studies we confirmed that Myf5 is directly regulated by Pax7 in myoblasts derived from satellite cells. Tandem affinity purification (TAP) and mass spectrometry were used to purify Pax7 together with its co-factors. This revealed that Pax7 associates with the Wdr5–Ash2L–MLL2 histone methyltransferase (HMT) complex that directs methylation of histone H3 lysine 4 (H3K4, refs 4–10). Binding of the Pax7–HMT complex to Myf5 resulted in H3K4 tri-methylation of surrounding chromatin. Thus, Pax7 induces chromatin modifications that stimulate transcriptional activation of target genes to regulate entry into the myogenic developmental programme.

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Figure 1: Candidate target genes are specifically activated by Pax7 in C2C12 myoblasts and are only weakly responsive to Pax3.
Figure 2: Identification of Pax7-interacting co-factors.
Figure 3: Pax7 immunocomplex has HMT activity and associates with sites of H3K4 methylation.
Figure 4: Pax7 regulates Myf5 expression directly in satellite cell-derived myoblasts.

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Acknowledgements

The authors are indebted to Doug Borris and Murray Smith for mass spectrometry analysis; Jeff Baker and Joyce Li for technical assistance; and to Marjorie Brand, Jennifer McCann, Mark Gillespie and Dave Picketts for critical input. This work was supported by grants to M.A.R. from the National Institutes of Health, the HHMI, the Canadian Institutes of Health Research, the Muscular Dystrophy Association and the CRC Program.

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Author notes

  1. Iain W. McKinnell and Jeff Ishibashi: These authors contributed equally to this work.

Authors and Affiliations

  1. Sprott Centre for Stem Cell Research, Ottawa Health Research Institute, 501 Smyth Road, Ottawa, K1H 8L6, Ontario, Canada

    Iain W. McKinnell, Jeff Ishibashi, Fabien Le Grand, Vincent G. J. Punch, Gregory C. Addicks, F. Jeffrey Dilworth & Michael A. Rudnicki

  2. Department of Medicine, University of Ottawa, 501 Smyth Road, Ottawa, K1H 8L6, Ontario, Canada

    Iain W. McKinnell, Jeff Ishibashi, Fabien Le Grand, Vincent G. J. Punch, Gregory C. Addicks, F. Jeffrey Dilworth & Michael A. Rudnicki

  3. Banting and Best Department of Medical Research, University of Toronto, 112 College Street, Ontario, M5G 1L6, Canada

    Jack F. Greenblatt

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Correspondence to Michael A. Rudnicki.

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McKinnell, I., Ishibashi, J., Le Grand, F. et al. Pax7 activates myogenic genes by recruitment of a histone methyltransferase complex. Nat Cell Biol 10, 77–84 (2008). https://doi.org/10.1038/ncb1671

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