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MBC in Press, published online ahead of print April 4, 2003
Mol. Biol. Cell 10.1091/mbc.E02-09-0600

A more recent version of this article appeared on July 1, 2003
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Submitted on September 18, 2002
Revised on February 26, 2003
Accepted on March 7, 2003

Requirement for down-regulation of the CCAAT-binding activity of the NF-Y transcription factor during skeletal muscle differentiation

Aymone Gurtner1, Isabella Manni1, Paola Fuschi1, Roberto Mantovani2, Fiorella Guadagni3, Ada Sacchi1, and Giulia Piaggio1*

1 Molecular Oncogenesis Laboratory, Experimental Oncology Department, Regina Elena Cancer Institute, Rome, Italy
2 Dipartimento di Biologia Animale, Università di Modena e Reggio, Modena, Italy
3 Laboratory of Clinical Pathology, Regina Elena Cancer Institute, Rome, Italy

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

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, while NF-YA alone is undetectable in terminally differentiated cells in vitro. By immuno-histochemistry 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 here 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.




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