Abstract
Previous works have established a unique function of MyoD in the control of muscle gene expression during DNA damage response in myoblasts. Phosphorylation by DNA damage-activated ABL tyrosine kinase transiently inhibits MyoD-dependent activation of transcription in response to genotoxic stress. We show here that ABL-MyoD signaling is also an essential component of the DNA repair machinery in myoblasts exposed to genotoxic stress. DNA damage promoted the recruitment of MyoD to phosphorylated Nbs1 (pNbs1)-containing repair foci, and this effect was abrogated by either ABL knockdown or the ABL kinase inhibitor imatinib. Upon DNA damage, MyoD and pNbs1 were detected on the chromatin to MyoD target genes without activating transcription. DNA damage-mediated tyrosine phosphorylation was required for MyoD recruitment to target genes, as the ABL phosphorylation-resistant MyoD mutant (MyoD Y30F) failed to bind the chromatin following DNA damage, while retaining the ability to activate transcription in response to differentiation signals. Moreover, MyoD Y30F exhibited an impaired ability to promote repair in a heterologous system, as compared with MyoD wild type (WT). Consistently, MyoD-null satellite cells (SCs) displayed impaired DNA repair that was rescued by reintroduction of MyoD WT but not by MyoD Y30F. In addition, inhibition of ABL kinase prevented MyoD WT-mediated rescue of DNA repair in MyoD-null SCs. These results identify an unprecedented contribution of MyoD to DNA repair and suggest that ABL-MyoD signaling coordinates DNA repair and transcription in myoblasts.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- DDR:
-
DNA damage response
- SCs:
-
Satellite cells
- GM:
-
growth medium
- DM:
-
differentiation medium
- MMS:
-
methyl methane sulfonate
- DOX:
-
doxorubicin
- MyHC:
-
myosin heavy chain
- MCK:
-
muscle creatine kinase
- Edu:
-
5-ethynyl-2′-deoxyuridine
- PLA:
-
proximity ligation assay
References
Dilworth FJ, Blais A . Epigenetic regulation of satellite cell activation during muscle regeneration. Stem Cell Res Ther 2011; 2: 18.
Lassar A, Munsterberg A . Wiring diagrams: regulatory circuits and the control of skeletal myogenesis. Curr Opin. Cell Biol 1994; 6: 432–442.
Tapscott SJ . The circuitry of a master switch: Myod and the regulation of skeletal muscle gene transcription. Development 2005; 132: 2685–2695.
Puri PL, Sartorelli V . Regulation of muscle regulatory factors by DNA-binding, interacting proteins, and post-transcriptional modifications. J Cell Physiol 2000; 185: 155–173.
Blais A, Tsikitis M, Acosta-Alvear D, Sharan R, Kluger Y, Dynlacht BD . An initial blueprint for myogenic differentiation. Genes Dev 2005; 19: 553–569.
Cao Y, Kumar RM, Penn BH, Berkes CA, Kooperberg C, Boyer LA et al. Global and gene-specific analyses show distinct roles for Myod and Myog at a common set of promoters. EMBO J 2006; 25: 502–511.
Cao Y, Yao Z, Sarkar D, Lawrence M, Sanchez GJ, Parker MH et al. Genome-wide MyoD binding in skeletal muscle cells: a potential for broad cellular reprogramming. Dev Cell 2010; 18: 662–674.
Kitzmann M, Carnac G, Vandromme M, Primig M, Lamb NJ, Fernandez A . The muscle regulatory factors MyoD and myf-5 undergo distinct cell cycle-specific expression in muscle cells. J Cell Biol 1998; 142: 1447–1459.
Zammit PS, Golding JP, Nagata Y, Hudon V, Partridge TA, Beauchamp JR . Muscle SCs adopt divergent fates: a mechanism for self-renewal? J Cell Biol 2004; 166: 347–357.
Megeney LA, Kablar B, Garrett K, Anderson JE, Rudnicki MA . MyoD is required for myogenic stem cell function in adult skeletal muscle. Genes Dev 1996; 10: 1173–1183.
Montarras D, Lindon C, Pinset C, Domeyne P . Cultured myf5 null and myoD null muscle precursor cells display distinct growth defects. Biol Cell 2000; 92: 565–572.
Zhang K, Sha J, Harter ML . Activation of Cdc6 by MyoD is associated with the expansion of quiescent myogenic SCs. J Cell Biol 2010; 188: 39–48.
Zhang K, Sha J, Harter ML . MyoD, a new function: ensuring "DNA licensing". Cell Cycle 2010; 9: 1871–1872.
Pallafacchina G, Francois S, Regnault B, Czarny B, Dive V, Cumano A et al. An adult tissue-specific stem cell in its niche: a gene profiling analysis of in vivo quiescent and activated muscle SCs. Stem Cell Res 2010; 4: 77–91.
Bartek J, Lukas C, Lukas J . Checking on DNA damage in S phase. Nat Rev Mol Cell Biol 2004; 5: 792–804.
Puri PL, Bhakta K, Wood LD, Costanzo A, Zhu J, Wang JY . A myogenic differentiation checkpoint activated by genotoxic stress. Nat Genet 2002; 32: 585–593.
Innocenzi A, Latella L, Messina G, Simonatto M, Marullo F, Berghella L et al. An evolutionarily acquired genotoxic response discriminates MyoD from Myf5, and differentially regulates hypaxial and epaxial myogenesis. EMBO R 2011; 12: 164–171.
Simonatto M, Giordani L, Marullo F, Minetti GC, Puri PL, Latella L . Coordination of cell cycle, DNA repair and muscle gene expression in myoblasts exposed to genotoxic stress. Cell Cycle 2011; 10: 2355–2363.
Simonatto M, Latella L, Puri PL . DNA damage and cellular differentiation: more questions than responses. J Cell Physiol 2007; 213: 642–648.
Xu Y, Price BD . Chromatin dynamics and the repair of DNA double strand breaks. Cell Cycle 2011; 10: 261–267.
Jackson SP, Bartek J . The DNA-damage response in human biology and disease. Nature 2009; 461: 1071–1078.
Allalou A, Wahlby C . BlobFinder, a tool for fluorescence microscopy image cytometry. Comput Methods Programs Biomed 2009; 94: 58–65.
Fredriksson S, Gullberg M, Jarvius J, Olsson C, Pietras K, Gustafsdottir SM et al. Protein detection using proximity-dependent DNA ligation assays. Nat Biotechnol 2002; 20: 473–477.
Asp P, Blum R, Vethantham V, Parisi F, Micsinai M, Cheng J et al. Genome-wide remodeling of the epigenetic landscape during myogenic differentiation. Proc Natl Acad Sci USA 2011; 108: E149–E158.
Guasconi V, Puri PL . Chromatin: the interface between extrinsic cues and the epigenetic regulation of muscle regeneration. Trends Cell Biol 2009; 19: 286–294.
Sartorelli V, Juan AH . Sculpting chromatin beyond the double helix: epigenetic control of skeletal myogenesis. Curr Topics Dev Biol 2011; 96: 57–83.
Puri PL, Sartorelli V, Yang XJ, Hamamori Y, Ogryzko VV, Howard BH et al. Differential roles of p300 and PCAF acetyltransferases in muscle differentiation. Mol Cell 1997; 1: 35–45.
Liu ZG, Baskaran R, Lea-Chou ET, Wood LD, Chen Y, Karin M et al. Three distinct signalling responses by murine fibroblasts to genotoxic stress. Nature 1996; 384: 273–276.
Wang JY, Ki SW . Choosing between growth arrest and apoptosis through the retinoblastoma tumour suppressor protein, Abl and p73. Biochem Soc Trans 2001; 29: 666–673.
Garinis GA, van der Horst GT, Vijg J, Hoeijmakers JH . DNA damage and ageing: new-age ideas for an age-old problem. Nat Cell Biol 2008; 10: 1241–1247.
Wang C, Jurk D, Maddick M, Nelson G, Martin-Ruiz C, von Zglinicki T . DDR and cellular senescence in tissues of aging mice. Aging Cell 2009; 8: 311–323.
Polesskaya A, Rudnicki MA . A MyoD-dependent differentiation checkpoint: ensuring genome integrity. Dev Cell 2002; 3: 757–758.
Frit P, Kwon K, Coin F, Auriol J, Dubaele S, Salles B et al. Transcriptional activators stimulate DNA repair. Mol Cell 2002; 10: 1391–1401.
Bell O, Tiwari VK, Thoma NH, Schubeler D . Determinants and dynamics of genome accessibility. Nat Rev Genet 2011; 12: 554–564.
Narciso L, Fortini P, Pajalunga D, Franchitto A, Liu P, Degan P et al. Terminally differentiated muscle cells are defective in base excision DNA repair and hypersensitive to oxygen injury. Proc Natl Acad Sci USA 2007; 104: 17010–17015.
Schmidt WM, Uddin MH, Dysek S, Moser-Thier K, Pirker C, Hoger H et al. DNA damage, somatic aneuploidy, and malignant sarcoma susceptibility in muscular dystrophies. PLoS Genet 2011; 7: e1002042.
Rosenblatt JD, Parry DJ, Partridge TA . Phenotype of adult mouse muscle myoblasts reflects their fiber type of origin. Differentiation 1996; 60: 39–45.
Furlan A, Stagni V, Hussain A, Richelme S, Conti F, Prodosmo A et al. Abl interconnects oncogenic Met and p53 core pathways in cancer cells. Cell Death Differ 2011; 18: 1608–1616.
Simone C, Forcales SV, Hill DA, Imbalzano AN, Latella L, Puri PL . p38 pathway targets SWI-SNF chromatin-remodeling complex to muscle-specific loci. Nat Genet 2004; 36: 738–743.
Forcales SV, Albini S, Giordani L, Malecova B, Cignolo L, Chernov A et al. Signal-dependent incorporation of MyoD-BAF60c into Brg1-based SWI/SNF chromatin-remodelling complex. EMBO J 2012; 31: 301–316.
Acknowledgements
PLP is a scientist at Sanford Children’ Health Center at Burnham and was partly supported by NIAMS (2R01AR052779-06). This work has benefited from research funding from the European Community’s Seventh Framework Programme in the project FP7-Health – 2009 ENDOSTEM 241440 (Activation of vasculature associated stem cells and muscle stem cells for the repair and maintenance of muscle tissue) and FILAS (to PLP) and Seventh Framework Programme-Myoage (Grant No. 223576), Fondazione Roma and Ateneo project (to AM). FM was supported by AFM PhD fellowship. LL was supported by a grant from Italian Foreign Ministry (MAE). We thank Dr. Michael Rudnicki for providing MyoD-null mice, Dr. Rossi for providing α7-integrin antibodies and Dr. Barila for providing Abl inhibitor and shRNA vectors.
Author Contributions
PLP designed and supervised the experiments of the manuscript. MS executed experiments of co-IF, ChIP and analysis of MyoD and ABL-null SCs. FM performed PLA and comet assays. LL performed PLA, IF during cell cycle in C2C12 myoblasts and IF of SCs during regeneration. JYJW provided ABL–CRE mice. FC and AM contributed to the image and data analysis. PLP conceived the experiments, wrote the manuscript and coordinated the revision of the manuscript. All authors discussed and commented the results of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Edited by G Melino
Supplementary Information accompanies this paper on Cell Death and Differentiation website
Rights and permissions
About this article
Cite this article
Simonatto, M., Marullo, F., Chiacchiera, F. et al. DNA damage-activated ABL-MyoD signaling contributes to DNA repair in skeletal myoblasts. Cell Death Differ 20, 1664–1674 (2013). https://doi.org/10.1038/cdd.2013.118
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/cdd.2013.118
Keywords
This article is cited by
-
DNA-PKcs regulates myogenesis in an Akt-dependent manner independent of induced DNA damage
Cell Death & Differentiation (2023)
-
SerpinE1 drives a cell-autonomous pathogenic signaling in Hutchinson–Gilford progeria syndrome
Cell Death & Disease (2022)
-
The Stat3-Fam3a axis promotes muscle stem cell myogenic lineage progression by inducing mitochondrial respiration
Nature Communications (2019)
-
Hsa-miR-520d-5p promotes survival in human dermal fibroblasts exposed to a lethal dose of UV irradiation
npj Aging and Mechanisms of Disease (2016)
-
Muscle gets stressed? p53 represses and protects
Cell Death & Differentiation (2015)
