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Myomaker is a membrane activator of myoblast fusion and muscle formation

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Abstract

Fusion of myoblasts is essential for the formation of multi-nucleated muscle fibres. However, the identity of muscle-specific proteins that directly govern this fusion process in mammals has remained elusive. Here we identify a muscle-specific membrane protein, named myomaker, that controls myoblast fusion. Myomaker is expressed on the cell surface of myoblasts during fusion and is downregulated thereafter. Overexpression of myomaker in myoblasts markedly enhances fusion, and genetic disruption of myomaker in mice causes perinatal death due to an absence of multi-nucleated muscle fibres. Remarkably, forced expression of myomaker in fibroblasts promotes fusion with myoblasts, demonstrating the direct participation of this protein in the fusion process. Pharmacological perturbation of the actin cytoskeleton abolishes the activity of myomaker, consistent with previous studies implicating actin dynamics in myoblast fusion. These findings reveal a long-sought myogenic fusion protein that controls mammalian myoblast fusion and provide new insights into the molecular underpinnings of muscle formation.

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Figure 1: Muscle-specific expression of myomaker.
Figure 2: Myomaker is essential for skeletal muscle development.
Figure 3: Control of myoblast fusion by myomaker.
Figure 4: Myomaker is expressed on the cell membrane of myoblasts.
Figure 5: Myomaker participates in the myoblast membrane fusion reaction.

References

  1. Chen, E. H. & Olson, E. N. Unveiling the mechanisms of cell-cell fusion. Science 308, 369–373 (2005)

    ADS  CAS  Article  Google Scholar 

  2. Bentzinger, C. F., Wang, Y. X. & Rudnicki, M. A. Building muscle: molecular regulation of myogenesis. Cold Spring Harb. Perspect. Biol. 4, a008342 (2012)

    Article  Google Scholar 

  3. Berkes, C. A. & Tapscott, S. J. MyoD and the transcriptional control of myogenesis. Semin. Cell Dev. Biol. 16, 585–595 (2005)

    CAS  Article  Google Scholar 

  4. Buckingham, M. Myogenic progenitor cells and skeletal myogenesis in vertebrates. Curr. Opin. Genet. Dev. 16, 525–532 (2006)

    CAS  Article  Google Scholar 

  5. Kang, J. S. & Krauss, R. S. Muscle stem cells in developmental and regenerative myogenesis. Curr. Opin. Clin. Nutr. Metab. Care 13, 243–248 (2010)

    CAS  Article  Google Scholar 

  6. Abmayr, S. M. & Pavlath, G. K. Myoblast fusion: lessons from flies and mice. Development 139, 641–656 (2012)

    CAS  Article  Google Scholar 

  7. Rochlin, K., Yu, S., Roy, S. & Baylies, M. K. Myoblast fusion: when it takes more to make one. Dev. Biol. 341, 66–83 (2010)

    CAS  Article  Google Scholar 

  8. Charrasse, S. et al. M-cadherin activates Rac1 GTPase through the Rho-GEF trio during myoblast fusion. Mol. Biol. Cell 18, 1734–1743 (2007)

    CAS  Article  Google Scholar 

  9. Charrasse, S., Meriane, M., Comunale, F., Blangy, A. & Gauthier-Rouviere, C. N-cadherin-dependent cell-cell contact regulates Rho GTPases and beta-catenin localization in mouse C2C12 myoblasts. J. Cell Biol. 158, 953–965 (2002)

    CAS  Article  Google Scholar 

  10. Schwander, M. et al. β1 integrins regulate myoblast fusion and sarcomere assembly. Dev. Cell 4, 673–685 (2003)

    CAS  Article  Google Scholar 

  11. Griffin, C. A., Kafadar, K. A. & Pavlath, G. K. MOR23 promotes muscle regeneration and regulates cell adhesion and migration. Dev. Cell 17, 649–661 (2009)

    CAS  Article  Google Scholar 

  12. Yagami-Hiromasa, T. et al. A metalloprotease-disintegrin participating in myoblast fusion. Nature 377, 652–656 (1995)

    ADS  CAS  Article  Google Scholar 

  13. Davis, R. L., Weintraub, H. & Lassar, A. B. Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51, 987–1000 (1987)

    CAS  Article  Google Scholar 

  14. Hasty, P. et al. Muscle deficiency and neonatal death in mice with a targeted mutation in the myogenin gene. Nature 364, 501–506 (1993)

    ADS  CAS  Article  Google Scholar 

  15. Vasyutina, E., Martarelli, B., Brakebusch, C., Wende, H. & Birchmeier, C. The small G-proteins Rac1 and Cdc42 are essential for myoblast fusion in the mouse. Proc. Natl Acad. Sci. USA 106, 8935–8940 (2009)

    ADS  CAS  Article  Google Scholar 

  16. Gruenbaum-Cohen, Y. et al. The actin regulator N-WASp is required for muscle-cell fusion in mice. Proc. Natl Acad. Sci. USA 109, 11211–11216 (2012)

    ADS  CAS  Article  Google Scholar 

  17. Wright, L. P. & Philips, M. R. Thematic review series: lipid posttranslational modifications. CAAX modification and membrane targeting of Ras. J. Lipid Res. 47, 883–891 (2006)

    CAS  Article  Google Scholar 

  18. Pei, J., Millay, D. P., Olson, E. N. & Grishin, N. V. CREST–a large and diverse superfamily of putative transmembrane hydrolases. Biol. Direct 6, 37 (2011)

    CAS  Article  Google Scholar 

  19. Corcoran, J. A. & Duncan, R. Reptilian reovirus utilizes a small type III protein with an external myristylated amino terminus to mediate cell-cell fusion. J. Virol. 78, 4342–4351 (2004)

    CAS  Article  Google Scholar 

  20. Chen, E. H. & Olson, E. N. Antisocial, an intracellular adaptor protein, is required for myoblast fusion in Drosophila. Dev. Cell 1, 705–715 (2001)

    CAS  Article  Google Scholar 

  21. Chen, E. H., Pryce, B. A., Tzeng, J. A., Gonzalez, G. A. & Olson, E. N. Control of myoblast fusion by a guanine nucleotide exchange factor, loner, and its effector ARF6. Cell 114, 751–762 (2003)

    CAS  Article  Google Scholar 

  22. Nowak, S. J., Nahirney, P. C., Hadjantonakis, A. K. & Baylies, M. K. Nap1-mediated actin remodeling is essential for mammalian myoblast fusion. J. Cell Sci. 122, 3282–3293 (2009)

    CAS  Article  Google Scholar 

  23. Laurin, M. et al. The atypical Rac activator Dock180 (Dock1) regulates myoblast fusion in vivo. Proc. Natl Acad. Sci. USA 105, 15446–15451 (2008)

    ADS  CAS  Article  Google Scholar 

  24. Powell, G. T. & Wright, G. J. Jamb and jamc are essential for vertebrate myocyte fusion. PLoS Biol. 9, e1001216 (2011)

    CAS  Article  Google Scholar 

  25. Oren-Suissa, M. & Podbilewicz, B. Cell fusion during development. Trends Cell Biol. 17, 537–546 (2007)

    CAS  Article  Google Scholar 

  26. Wilson, N. F. & Snell, W. J. Microvilli and cell-cell fusion during fertilization. Trends Cell Biol. 8, 93–96 (1998)

    CAS  Article  Google Scholar 

  27. Shilagardi, K. et al. Actin-propelled invasive membrane protrusions promote fusogenic protein engagement during cell-cell fusion. Science 340, 359–363 (2013)

    ADS  CAS  Article  Google Scholar 

  28. Skarnes, W. C. et al. A conditional knockout resource for the genome-wide study of mouse gene function. Nature 474, 337–342 (2011)

    CAS  Article  Google Scholar 

  29. Hargrave, M. & Koopman, P. In situ hybridization of whole-mount embryos. Methods Mol. Biol. 123, 279–289 (2000)

    CAS  PubMed  Google Scholar 

  30. Kitamura, T. et al. Efficient screening of retroviral cDNA expression libraries. Proc. Natl Acad. Sci. USA 92, 9146–9150 (1995)

    ADS  CAS  Article  Google Scholar 

  31. Song, K. et al. Heart repair by reprogramming non-myocytes with cardiac transcription factors. Nature 485, 599–604 (2012)

    ADS  CAS  Article  Google Scholar 

  32. Shelton, J. M., Lee, M. H., Richardson, J. A. & Patel, S. B. Microsomal triglyceride transfer protein expression during mouse development. J. Lipid Res. 41, 532–537 (2000)

    CAS  PubMed  Google Scholar 

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Acknowledgements

We thank J. Cabrera for graphics, the Transgenic Technology Center at University of Texas Southwestern Medical Center for embryonic stem cell injections, J.A. Richardson for histology and the University of Texas Southwestern Live Cell Imaging Facility (K. Luby-Phelps and A. Budge) for microscopy and live cell-imaging assistance. We thank D. Rosenbaum, W. Snell, S. Schmid, J. Seemann and members of the Olson laboratory for scientific discussions. D.P.M. was financed by a National Institutes of Health National Research Service Award Fellowship (F32AR05948403). This work was supported by grants from the National Institutes of Health (HL-077439, HL-111665, HL093039 and U01-HL-100401) and the Robert A. Welch Foundation (grant 1-0025) (to E.N.O.).

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Contributions

D.P.M. and E.N.O. conceived the project and designed the experiments. D.P.M., J.R.O., L.B.S., S.B. and J.M.S. performed experiments. D.P.M. and R.B.-D. wrote the animal protocol. D.P.M. and E.N.O. analysed the data and prepared the manuscript.

Corresponding author

Correspondence to Eric N. Olson.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-8. (PDF 9769 kb)

Time-lapse microscopy of C2C12 myoblasts infected with GFP and Myomaker

C2C12 cells were infected with GFP and Myomaker retrovirus and induced to differentiate. This video depicts a myoblast fusing to a myoblast (arrowhead) and fusion of two myotubes (arrows). Myomaker not only increases myoblast-myotube fusion, but also myotube-myotube fusion. The video represents 32 hours of culture, beginning 48 hours after differentiation. Frames are 15 minutes apart and video speed is 3 frames/second. (MOV 11306 kb)

Myomaker-infected fibroblasts fusing to muscle cells

10T½ fibroblasts were infected with GFP and Myomaker. C2C12 myoblasts were infected with dsRed and then mixed with the GFP+Myomaker-infected fibroblasts and induced to differentiate. Red/Green chimeric myotubes, generated from the fusion of myoblasts and fibroblasts, can be visualized fusing with a C2C12 cell (red arrow) and fibroblasts (green arrows). The video represents 40 hours of culture, beginning 48 hours after differentiation. Frames are 15 minutes apart and video speed is 3 frames/second. (MOV 27854 kb)

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Millay, D., O’Rourke, J., Sutherland, L. et al. Myomaker is a membrane activator of myoblast fusion and muscle formation. Nature 499, 301–305 (2013). https://doi.org/10.1038/nature12343

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