Dephosphorylation of HDAC4 by PP2A-Bdelta unravels a new role for the HDAC4/MEF2 axis in myoblast fusion.

[en] Muscle formation is controlled by a number of key myogenic transcriptional regulators that govern stage-specific gene expression programs and act as terminal effectors of intracellular signaling pathways. To date, the role of phosphatases in the signaling cascades instructing muscle development remains poorly understood. Here, we show that a specific PP2A-B55delta holoenzyme is necessary for skeletal myogenesis. The primary role of PP2A-B55delta is to dephosphorylate histone deacetylase 4 (HDAC4) following myocyte differentiation and ensure repression of Myocyte enhancer factor 2D (MEF2D)-dependent gene expression programs during myogenic fusion. As a crucial HDAC4/MEF2D target gene that governs myocyte fusion, we identify ArgBP2, an upstream inhibitor of Abl, which itself is a repressor of CrkII signaling. Consequently, cells lacking PP2A-B55delta show upregulation of ArgBP2 and hyperactivation of CrkII downstream effectors, including Rac1 and FAK, precluding cytoskeletal and membrane rearrangements associated with myoblast fusion. Both in vitro and in zebrafish, loss-of-function of PP2A-B55delta severely impairs fusion of myocytes and formation of multinucleated muscle fibers, without affecting myoblast differentiation. Taken together, our results establish PP2A-B55delta as the first protein phosphatase to be involved in myoblast fusion and suggest that reversible phosphorylation of HDAC4 may coordinate differentiation and fusion events during myogenesis.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Veloso, Alexandra

Martin, Maud

Bruyr, Jonathan

O'Grady, Tina ; Université de Liège - ULiège > Molecular Biology of Diseases-Gene Expression & Cancer

Deroanne, Christophe ; Université de Liège - ULiège > Cancer-Connective Tissue Biology

Mottet, Denis ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Département des sciences biomédicales et précliniques

Twizere, Jean-Claude ; Université de Liège - ULiège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Molecular Biology of Diseases-Viral Interactomes Network

Cherrier, Thomas ^✱

Dequiedt, Franck ^✱; Université de Liège - ULiège > Département des sciences de la vie > Génétique et biologie moléculaires animales

^✱ These authors have contributed equally to this work.

Language :

English

Title :

Dephosphorylation of HDAC4 by PP2A-Bdelta unravels a new role for the HDAC4/MEF2 axis in myoblast fusion.

Publication date :

2019

Journal title :

Cell death & disease

eISSN :

2041-4889

Volume :

Issue :

Pages :

512

Peer reviewed :

Peer reviewed

Available on ORBi :

since 08 August 2019

Statistics

Number of views

210 (54 by ULiège)

Number of downloads

73 (8 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

Hindi, S. M., Tajrishi, M. M. & Kumar, A. Signaling mechanisms in mammalian myoblast fusion. Sci Signal 6, re2 (2013).
Buckingham, M. & Rigby, P. W. Gene regulatory networks and transcriptional mechanisms that control myogenesis. Dev Cell 28, 225–238 (2014).
Taylor, M. V. & Hughes, S. M. Mef2 and the skeletal muscle differentiation program. Semin Cell Dev Biol 72, 33–44 (2017).
Lu, J., McKinsey, T. A., Zhang, C. L. & Olson, E. N. Regulation of skeletal myogenesis by association of the MEF2 transcription factor with class II histone deacetylases. Mol Cell 6, 233–244 (2000).
Dressel, U. et al. A dynamic role for HDAC7 in MEF2-mediated muscle differentiation. J Biol Chem 276, 17007–17013 (2001).
Gao, C., Liu, Y., Lam, M. & Kao, H. Y. Histone deacetylase 7 (HDAC7) regulates myocyte migration and differentiation. Biochim Biophys Acta 1803, 1186–1197 (2010).
Haberland, M. et al. Regulation of HDAC9 gene expression by MEF2 establishes a negative-feedback loop in the transcriptional circuitry of muscle differentiation. Mol Cell Biol 27, 518–525 (2007).
Zhang, C. L., McKinsey, T. A. & Olson, E. N. The transcriptional corepressor MITR is a signal-responsive inhibitor of myogenesis. Proc Natl Acad Sci USA 98, 7354–7359 (2001).
Martin, M., Kettmann, R. & Dequiedt, F. Class IIa histone deacetylases: regulating the regulators. Oncogene 26, 5450–5467 (2007).
McKinsey, T. A., Zhang, C. L. & Olson, E. N. Signaling chromatin to make muscle. Curr Opin Cell Biol 14, 763–772 (2002).
Knight, J. D. & Kothary, R. The myogenic kinome: protein kinases critical to mammalian skeletal myogenesis. Skelet Muscle 1, 29 (2011).
Ciccone, M., Calin, G. A. & Perrotti, D. From the biology of PP2A to the PADs for therapy of hematologic malignancies. Front Oncol 5, 21 (2015).
Hunt, T. On the regulation of protein phosphatase 2A and its role in controlling entry into and exit from mitosis. Adv Biol Regul 53, 173–178 (2013).
Janssens, V. & Rebollo, A. The role and therapeutic potential of Ser/Thr phosphatase PP2A in apoptotic signalling networks in human cancer cells. Curr Mol Med 12, 268–287 (2012).
Sents, W., Ivanova, E., Lambrecht, C., Haesen, D. & Janssens, V. The biogenesis of active protein phosphatase 2A holoenzymes: a tightly regulated process creating phosphatase specificity. FEBS J 280, 644–661 (2013).
Emi, N., Friedmann, T. & Yee, J. K. Pseudotype formation of murine leukemia virus with the G protein of vesicular stomatitis virus. J Virol 65, 1202–1207 (1991).
Deroanne, C., Vouret-Craviari, V., Wang, B. & Pouyssegur, J. EphrinA1 inactivates integrin-mediated vascular smooth muscle cell spreading via the Rac/PAK pathway. J Cell Sci 116, 1367–1376 (2003).
Patro, R., Duggal, G., Love, M. I., Irizarry, R. A. & Kingsford, C. Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods 14, 417–419 (2017).
Soneson, C., Love, M. I. & Robinson, M. D. Differential analyses for RNA-seq: transcript-level estimates improve gene-level inferences. F1000Res 4, 1521 (2015).
Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15, 550 (2014).
Huang da, W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4, 44–57 (2009).
Wales, S., Hashemi, S., Blais, A. & McDermott, J. C. Global MEF2 target gene analysis in cardiac and skeletal muscle reveals novel regulation of DUSP6 by p38MAPK-MEF2 signaling. Nucleic Acids Res 42, 11349–11362 (2014).
Swingle, M., Ni, L. & Honkanen, R. E. Small-molecule inhibitors of ser/thr protein phosphatases: specificity, use and common forms of abuse. Methods Mol Biol 365, 23–38 (2007).
Kong, M., Ditsworth, D., Lindsten, T. & Thompson, C. B. Alpha4 is an essential regulator of PP2A phosphatase activity. Mol Cell 36, 51–60 (2009).
Li, M., Guo, H. & Damuni, Z. Purification and characterization of two potent heat-stable protein inhibitors of protein phosphatase 2A from bovine kidney. Biochemistry 34, 1988–1996 (1995).
Melnikova, I. N., Bounpheng, M., Schatteman, G. C., Gilliam, D. & Christy, B. A. Differential biological activities of mammalian Id proteins in muscle cells. Exp Cell Res 247, 94–104 (1999).
Molkentin, J. D. & Olson, E. N. Defining the regulatory networks for muscle development. Curr Opin Genet Dev 6, 445–453 (1996).
Sohn, R. L. et al. A role for nephrin, a renal protein, in vertebrate skeletal muscle cell fusion. Proc Natl Acad Sci USA 106, 9274–9279 (2009).
Swailes, N. T., Colegrave, M., Knight, P. J. & Peckham, M. Non-muscle myosins 2A and 2B drive changes in cell morphology that occur as myoblasts align and fuse. J Cell Sci 119, 3561–3570 (2006).
Peckham, M. Engineering a multi-nucleated myotube, the role of the actin cytoskeleton. J Microsc 231, 486–493 (2008).
Wei, L. et al. RhoA signaling via serum response factor plays an obligatory role in myogenic differentiation. J Biol Chem 273, 30287–30294 (1998).
Sanders, L. C., Matsumura, F., Bokoch, G. M. & de Lanerolle, P. Inhibition of myosin light chain kinase by p21-activated kinase. Science 283, 2083–2085 (1999).
Ridley, A. J. Life at the leading edge. Cell 145, 1012–1022 (2011).
Delorme, V. et al. Cofilin activity downstream of Pak1 regulates cell protrusion efficiency by organizing lamellipodium and lamella actin networks. Dev Cell 13, 646–662 (2007).
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).
Brugnera, E. et al. Unconventional Rac-GEF activity is mediated through the Dock180-ELMO complex. Nat Cell Biol 4, 574–582 (2002).
Birge, R. B., Kalodimos, C., Inagaki, F. & Tanaka, S. Crk and CrkL adaptor proteins: networks for physiological and pathological signaling. Cell Commun Signal 7, 13 (2009).
Zvara, A. et al. Activation of the focal adhesion kinase signaling pathway by structural alterations in the carboxyl-terminal region of c-Crk II. Oncogene 20, 951–961 (2001).
Clemente, C. F., Corat, M. A., Saad, S. T. & Franchini, K. G. Differentiation of C2C12 myoblasts is critically regulated by FAK signaling. Am J Physiol Regul Integr Comp Physiol 289, R862–R870 (2005).
Quach, N. L., Biressi, S., Reichardt, L. F., Keller, C. & Rando, T. A. Focal adhesion kinase signaling regulates the expression of caveolin 3 and beta1 integrin, genes essential for normal myoblast fusion. Mol Biol Cell 20, 3422–3435 (2009).
Martin, M. et al. PP2A regulatory subunit Balpha controls endothelial contractility and vessel lumen integrity via regulation of HDAC7. EMBO J 32, 2491–2503 (2013).
Martin, M., Kettmann, R. & Dequiedt, F. Class IIa histone deacetylases: conducting development and differentiation. Int J Dev Biol 53, 291–301 (2009).
Potthoff, M. J. & Olson, E. N. MEF2: a central regulator of diverse developmental programs. Development 134, 4131–4140 (2007).
Soubeyran, P., Barac, A., Szymkiewicz, I. & Dikic, I. Cbl-ArgBP2 complex mediates ubiquitination and degradation of c-Abl. Biochem J 370, 29–34 (2003).
Sampath, S. C., Sampath, S. C. & Millay, D. P. Myoblast fusion confusion: the resolution begins. Skelet Muscle 8, 3 (2018).
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).
Kim, J. H., Jin, P., Duan, R. & Chen, E. H. Mechanisms of myoblast fusion during muscle development. Curr Opin Genet Dev 32, 162–170 (2015).
Bassel-Duby, R. & Olson, E. N. Signaling pathways in skeletal muscle remodeling. Annu Rev Biochem 75, 19–37 (2006).
Lu, J., McKinsey, T. A., Nicol, R. L. & Olson, E. N. Signal-dependent activation of the MEF2 transcription factor by dissociation from histone deacetylases. Proc Natl Acad Sci USA 97, 4070–4075 (2000).
Cadot, B. et al. Loss of histone deacetylase 4 causes segregation defects during mitosis of p53-deficient human tumor cells. Cancer Res 69, 6074–6082 (2009).
Paroni, G. et al. PP2A regulates HDAC4 nuclear import. Mol Biol Cell 19, 655–667 (2008).
Parra, M. Class IIa HDACs - new insights into their functions in physiology and pathology. FEBS J 282, 1736–1744 (2015).