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Molecular and Cellular Biology, April 2003, p. 2893-2906, Vol. 23, No. 8
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.8.2893-2906.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

MyoD Stimulates RB Promoter Activity via the CREB/p300 Nuclear Transduction Pathway{dagger}

Alessandra Magenta, Carlo Cenciarelli, Francesca De Santa, Paola Fuschi,{ddagger} Fabio Martelli,§ Maurizia Caruso,* and Armando Felsani*

CNR, Istituto Neurobiologia e Medicina Molecolare, I-00137 Rome, Italy

Received 6 June 2002/ Returned for modification 25 July 2002/ Accepted 15 January 2003

The induction of RB gene transcription by MyoD is a key event in the process of skeletal muscle differentiation, because elevated levels of the retinoblastoma protein are essential for myoblast cell cycle arrest as well as for the terminal differentiation and survival of postmitotic myocytes. We previously showed that MyoD stimulates transcription from the RB promoter independently of direct binding to promoter sequences. Here we demonstrate that stimulation by MyoD requires a cyclic AMP-responsive element (CRE) in the RB promoter, bound by the transcription factor CREB in differentiating myoblasts. We also show that both the CREB protein level and the level of phosphorylation of the CREB protein at Ser-133 rapidly increase at the onset of muscle differentiation and that both remain high throughout the myogenic process. Biochemical and functional evidence indicates that in differentiating myoblasts, MyoD becomes associated with CREB and is targeted to the RB promoter CRE in a complex also containing the p300 transcriptional coactivator. The resulting multiprotein complex stimulates transcription from the RB promoter. These and other observations strongly suggest that MyoD functions by promoting the efficient recruitment of p300 by promoter-bound, phosphorylated CREB.

* Corresponding author. Mailing address for Maurizia Caruso: CNR, Istituto Neurobiologia e Medicina Molecolare, Viale Marx 43, 00137 Rome, Italy. Phone: 39 06 86090303. Fax: 39 06 86090325. E-mail: maurizia.caruso{at}inemm.cnr.it. Mailing address for Armando Felsani: CNR, Istituto Neurobiologia e Medicina Molecolare, Viale Marx 43, 00137 Rome, Italy. Phone: 39 06 86090500. Fax: 39 06 86090325. E-mail: felsani{at}inemm.cnr.it.

{dagger} This work is dedicated to the memory of Franco Tatò.

{ddagger} Present address: Laboratorio di Oncogenesi Molecolare, Dipartimento di Oncologia Sperimentale, Istituto Regina Elena, 00158 Rome, Italy.

§ Present address: Laboratorio Patologia Vascolare, IRCCS, Istituto Dermopatico dell'Immacolata, 00167 Rome, Italy.

Molecular and Cellular Biology, April 2003, p. 2893-2906, Vol. 23, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.8.2893-2906.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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