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Vol. 14, Issue 7, 2706-2715, July 2003

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Requirement for Down-Regulation of the CCAAT-binding Activity of the NF-Y Transcription Factor during Skeletal Muscle Differentiation

Aymone Gurtner ^*, Isabella Manni ^*, Paola Fuschi ^*, Roberto Mantovani , Fiorella Guadagni , Ada Sacchi ^*, and Giulia Piaggio ^* 

^*Molecular Oncogenesis Laboratory, Experimental Oncology Department, Regina Elena Cancer Institute, Rome, Italy; Laboratory of Clinical Pathology, Regina Elena Cancer Institute, Rome, Italy; and Dipartimento di Biologia Animale, Università di Modena e Reggio, Modena, Italy

Submitted September 18, 2002; Revised February 26, 2003; Accepted March 7, 2003
Monitoring Editor: Marvin P. Wickens

NF-Y is composed of three subunits, NF-YA, NF-YB, and NF-YC, all required for DNA binding. All subunits are expressed in proliferating skeletal muscle cells, whereas NF-YA alone is undetectable in terminally differentiated cells in vitro. By immunohistochemistry, we show that the NF-YA protein is not expressed in the nuclei of skeletal and cardiac muscle cells in vivo. By chromatin immunoprecipitation experiments, we demonstrate herein that NF-Y does not bind to the CCAAT boxes of target promoters in differentiated muscle cells. Consistent with this, the activity of these promoters is down-regulated in differentiated muscle cells. Finally, forced expression of the NF-YA protein in cells committed to differentiate leads to an impairment in the down-regulation of cyclin A, cyclin B1, and cdk1 expression and is accompanied by a delay in myogenin expression. Thus, our results indicate that the suppression of NF-Y function is of crucial importance for the inhibition of several cell cycle genes and the induction of the early muscle-specific program in postmitotic muscle cells.

Corresponding author. E-mail address: piaggio{at}ifo.it.

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