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A myogenic differentiation checkpoint activated by genotoxic stress

Nature Genetics volume 32, pages 585–593 (2002)Cite this article

Abstract

Cell-cycle checkpoints help to protect the genomes of proliferating cells under genotoxic stress. In multicellular organisms, cell proliferation is often directed toward differentiation during development and throughout adult homeostasis. To prevent the formation of differentiated cells with genetic instability, we hypothesized that genotoxic stress may trigger a differentiation checkpoint. Here we show that exposure to genotoxic agents causes a reversible inhibition of myogenic differentiation. Muscle-specific gene expression is suppressed by DNA-damaging agents if applied prior to differentiation induction but not after the differentiation program is established. The myogenic determination factor, MyoD (encoded by Myod1), is a target of the differentiation checkpoint in myoblasts. The inhibition of MyoD by DNA damage requires a functional c-Abl tyrosine kinase (encoded by Abl1), but occurs in cells deficient for p53 (transformation-related protein 53, encoded by Trp53) or c-Jun (encoded by the oncogene Jun). These results support the idea that genotoxic stress can regulate differentiation, and identify a new biological function for DNA damage–activated signaling network.

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Figure 1: Inhibition of myogenic differentiation by genotoxic stress.
Figure 2: Genotoxic stress inhibits MyoD function.
Figure 3: Inhibition of myogenic conversion by genotoxic drugs requires c-Abl.
Figure 4: Ectopic expression of nuclear c-Abl sensitizes myoblasts to an MMS-induced differentiation checkpoint.
Figure 5: Tyrosine phosphorylation of MyoD by c-Abl.
Figure 6: Tyrosine-30 phosphorylation of MyoD correlates with inhibition of its transactivation function.

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Acknowledgements

We thank V. Sartorelli for discussion and for providing retroviral MyoD and MyoD (Tyr30Phe); M. Montminy, P.J. Woodring and M. Levrero for stimulating discussions; T. Hunter for critically reading the manuscript; L. Kedes and C. Poizat for providing adenoviral FLAG–MyoD; B. Paterson for providing His–MyoD; K. Du for providing polyclonal 293 cells stably transfected with Gal4–Luc; P. Stiegler for C2C12 cells with integrated luciferase reporters (Ckmm–Luc or Trp53–Luc); I.C. Hunton in the laboratory of J.Y.J.W. for the generation of MEFs and 3T3 cells; and J. Meisenhelder and U. Jhala for technical advice on phosphopeptide mapping. This work was supported by the American Cancer Society Postdoctoral Fellowship (L.D.W.), Human Frontiers Postdoctoral Fellowship (P.L.P.), Muscular Dystrophy Association (P.L.P.) and the US National Cancer Institute (J.Y.J.W.). P.L.P. is currently an Assistant Scientist of the Dulbecco Telethon Institute.

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Authors and Affiliations

  1. Dulbecco Telethon Institute at Laboratory of Gene Expression, Fondazione Andrea Cesalpino University of Rome “La Sapienza”, Rome, 00161, Italy

    Pier Lorenzo Puri & Antonio Costanzo

  2. Division of Biological Sciences and the Cancer Center, University of California, San Diego, La Jolla, 92093-0322, California, USA

    Pier Lorenzo Puri, Kunjan Bhakta, Lauren D. Wood, Jiangyu Zhu & Jean Y. J. Wang

  3. Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, 92037-1099, California, USA

    Pier Lorenzo Puri

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  1. Pier Lorenzo Puri
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Correspondence to Pier Lorenzo Puri or Jean Y. J. Wang.

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Puri, P., Bhakta, K., Wood, L. et al. A myogenic differentiation checkpoint activated by genotoxic stress. Nat Genet 32, 585–593 (2002). https://doi.org/10.1038/ng1023

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