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Molecular and Cellular Biology, November 2001, p. 7761-7774, Vol. 21, No. 22
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.22.7761-7774.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

p300 Acts as a Transcriptional Coactivator for Mammalian Notch-1

Franz Oswald,1 Birgitt Täuber,2 Thomas Dobner,2 Soizic Bourteele,1 Ulrike Kostezka,1 Guido Adler,1 Susanne Liptay,3 and Roland M. Schmid1,*

Departments of Internal Medicine1 and Pediatrics,3 University of Ulm, D-89081 Ulm, and Institute for Medical Microbiology and Hygiene, University of Regensburg, D-93053 Regensburg,2 Germany

Received 16 March 2001/Returned for modification 29 April 2001/Accepted 2 August 2001

Notch-1 belongs to a family of transmembrane receptor proteins that direct the decisions as to various cell fates. After ligand binding, a proteolytic cleavage step occurs and the intracellular part of Notch-1, Notch-1-IC, translocates into the nucleus, where it targets the DNA binding protein RBP-Jkappa /CBF1. RBP-Jkappa mediates repression through recruitment of a histone deacetylase-containing complex. The Notch-1-IC/RBP-Jkappa complex overcomes repression and activates the transcription of Notch target genes. We have identified a novel domain in Notch-1-IC, the EP domain, which is indispensable for full transcriptional activation. This transactivation domain is localized adjacent to the ankyrin repeats of Notch-1-IC. In cotransfection experiments, Notch-1-IC-mediated transcriptional activation was inhibited by E1A12S and p53, two proteins, which interfere with the function of the common coactivator p300. Protein-protein interaction assays demonstrated the association of Notch-1-IC and the CH3 region of p300. In addition, the interaction of mammalian Notch-1-IC with p300 was destabilized after deletion of the EP domain of Notch-1-IC. Based on physical interaction with Notch-1-IC and coactivator functions of p300, we propose a model for Notch-1-mediated gene regulation via p300.

* Corresponding author. Mailing address: Department of Internal Medicine I, Robert-Koch-Strasse 8, D-89081 Ulm, Germany. Phone: 49-731-50-24305. Fax: 49-731-50-24302. E-mail: roland.schmid{at}medizin.uni-ulm.de.

Molecular and Cellular Biology, November 2001, p. 7761-7774, Vol. 21, No. 22
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.22.7761-7774.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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