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Molecular and Cellular Biology, July 1999, p. 5073-5082, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Cell Cycle Withdrawal Promotes Myogenic Induction of Akt, a Positive Modulator of Myocyte Survival

Yasushi Fujio,1 Kun Guo,1,2 Toshiaki Mano,1 Yasuhiro Mitsuuchi,3 Joseph R. Testa,3 and Kenneth Walsh1,4,*

Division of Cardiovascular Research, St. Elizabeth's Medical Center of Boston, Tufts University School of Medicine, Boston, Massachusetts 021351; Department of Cardiovascular Diseases, Rhone-Poulenc Rorer, Inc., Collegeville, Pennsylvania 194262; Department of Medical Oncology, Fox Chase Cancer Center, Philadelphia, Pennsylvania 191113; and Program in Cell, Molecular and Developmental Biology, Sackler School of Biomedical Studies, Tufts University School of Medicine, Boston, Massachusetts 021114

Received 25 November 1998/Returned for modification 11 January 1999/Accepted 7 April 1999

During myogenesis, proliferating myoblasts withdraw from the cell cycle, acquire an apoptosis-resistant phenotype, and differentiate into myotubes. Previous studies indicate that myogenic induction of the cyclin-dependent kinase inhibitor p21 results in an inhibition of apoptotic cell death in addition to its role as a negative cell cycle regulator. Here we demonstrate that the protein encoded by the Akt proto-oncogene is induced in C2C12 cells during myogenic differentiation with a corresponding increase in kinase activity. In differentiating cultures, expression of dominant-negative forms of Akt increase the frequency of cell death whereas expression of wild-type Akt protects against death, indicating that Akt is a positive modulator of myocyte survival. Antisense oligonucleotides against p21 block cell cycle withdrawal, inhibit Akt induction, and enhance cell death in differentiating myocyte cultures. Adenovirus-mediated transfer of wild-type or constitutively active Akt constructs confer partial resistance to cell death under conditions where cell cycle exit is blocked by the antisense oligonucleotides. Collectively, these data indicate that cell cycle withdrawal facilitates the induction of Akt during myogenesis, promoting myocyte survival.

* Corresponding author. Mailing address: Division of Cardiovascular Research, St. Elizabeth's Medical Center, 736 Cambridge St., Boston, MA 02135. Phone: (617) 562-7501. Fax: (617) 562-7506. E-mail: kwalsh{at}opal.tufts.edu.

Molecular and Cellular Biology, July 1999, p. 5073-5082, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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