NDLI: Synthesis of Mitochondrial DNA Precursors during Myogenesis, an Analysis in Purified C2C12 $Myotubes^{*}$

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Synthesis of Mitochondrial DNA Precursors during Myogenesis, an Analysis in Purified C2C12 $Myotubes^{*}$

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Synthesis of Mitochondrial DNA Precursors during Myogenesis, an Analysis in Purified C2C12 $Myotubes^{*}$

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Chemello, Francesco Romualdi, Chiara Rampazzo, Chiara Franzolin, Elisa Laveder, Paolo Bianchi, Vera Frangini, Miriam
Description	Author Affiliation: Frangini M ( Department of Biology, University of Padova, 35131 Padova, Italy.)
Abstract	During myogenesis, myoblasts fuse into multinucleated myotubes that acquire the contractile fibrils and accessory structures typical of striated skeletal muscle fibers. To support the high energy requirements of muscle contraction, myogenesis entails an increase in mitochondrial (mt) mass with stimulation of mtDNA synthesis and consumption of DNA precursors (dNTPs). Myotubes are quiescent cells and as such down-regulate dNTP production despite a high demand for dNTPs. Although myogenesis has been studied extensively, changes in dNTP metabolism have not been examined specifically. In differentiating cultures of C2C12 myoblasts and purified myotubes, we analyzed expression and activities of enzymes of dNTP biosynthesis, dNTP pools, and the expansion of mtDNA. Myotubes exibited pronounced post-mitotic modifications of dNTP synthesis with a particularly marked down-regulation of de novo thymidylate synthesis. Expression profiling revealed the same pattern of enzyme down-regulation in adult murine muscles. The mtDNA increased steadily after myoblast fusion, turning over rapidly, as revealed after treatment with ethidium bromide. We individually down-regulated p53R2 ribonucleotide reductase, thymidine kinase 2, and deoxyguanosine kinase by siRNA transfection to examine how a further reduction of these synthetic enzymes impacted myotube development. Silencing of p53R2 had little effect, but silencing of either mt kinase caused 50% mtDNA depletion and an unexpected decrease of all four dNTP pools independently of the kinase specificity. We suggest that during development of myotubes the shortage of even a single dNTP may affect all four pools through dysregulation of ribonucleotide reduction and/or dissipation of the non-limiting dNTPs during unproductive elongation of new DNA chains.
ISSN	00219258
e-ISSN	1083351X
Journal	Journal of Biological Chemistry
Issue Number	8
Volume Number	288
Language	English
Publisher	American Society for Biochemistry and Molecular Biology (United States)
Publisher Date	2013-02-22
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA, Mitochondrial Genetics Muscle Development Physiology Animals Cell Differentiation Cell Line Cytosol Metabolism Gene Silencing Mice Mitochondria Mitochondrial Diseases Models, Biological Muscle, Skeletal Nucleic Acid Hybridization Nucleotides Phosphotransferases (Alcohol Group Acceptor) RNA, Small Interfering Ribonucleotides Research Support, Non-U.S. Gov't Biochemistry Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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