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J Biol Chem, Vol. 273, Issue 14, 8492-8501, April 3, 1998

MyoD Is Indispensable for Muscle-specific Alternative Splicing in Mouse Mitochondrial ATP Synthase -Subunit Pre-mRNA

Masaru Ichida^§, Hitoshi Endo, Uichi Ikeda^§, Chie Matsuda, Eriko Ueno, Kazuyuki Shimada^§, and Yasuo Kagawa

From the Departments of  Biochemistry and ^§ Cardiology, Jichi Medical School, Minamikawachi-machi, Kawachi-gun, Tochigi 329-04, Japan

Muscle-specific alternative RNA splicing is an essential step during myogenesis. In this paper, we report that a muscle-specific transcription factor, MyoD, plays a central role in the induction of muscle-specific alternative splicing during myogenesis. Recently, we reported that muscle and nonmuscle isoforms of the mitochondrial ATP synthase -subunit (F₁) were generated by alternative splicing and that acidic stimulation promoted this muscle-specific alternative splicing (Endo, H., Matsuda, C., and Kagawa, Y. (1994) J. Biol. Chem. 269, 12488-12493). In this report, mouse myoblasts are shown to express the muscle-specific isoform of F₁ after induction with low-serum medium (differentiation medium) or acidic medium, although myotube formation was not detected after acidic induction. RNA blot analysis revealed that the expression levels of both MEF2 and myogenin were increased by low-serum induction, but not by acidic induction. High expression of MyoD mRNA was observed after both types of induction. Overexpression of exogenous MyoD in fibroblasts showed that MyoD was necessary for muscle-specific alternative splicing in both types of induction. Exogenous Id, a negative regulator of MyoD, blocked muscle-specific alternative splicing of F₁ pre-mRNA by both types of induction. In addition, MyoD induced several muscle-specific alternative splicings, including structural protein pre-mRNAs such as -tropomyosin and neural-cell adhesion molecule and transcriptional protein pre-mRNAs such as MEF2A and MEF2D. Our analysis of the two induction systems shows a common MyoD-dependent mechanism of muscle-specific alternative splicing in several genes, independent of MEF2 and myogenin.

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