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Molecular and Cellular Biology, November 2002, p. 7812-7819, Vol. 22, No. 22
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.22.7812-7819.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

p300 and PCAF Act Cooperatively To Mediate Transcriptional Activation from Chromatin Templates by Notch Intracellular Domains In Vitro

Annika E. Wallberg,1* Kia Pedersen,2 Urban Lendahl,2 and Robert G. Roeder1

Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, New York, New York 10021,1 Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institutet, SE-17177, Stockholm, Sweden2

Received 24 May 2002/ Returned for modification 6 August 2002/ Accepted 13 August 2002

Ligand activation of Notch receptors leads to release of the intracellular receptor domain (Notch IC), which translocates to the nucleus and interacts with the DNA-binding protein RBP-J{kappa} to control expression of specific target genes. A number of proteins have been shown to interact with Notch ICs and to modulate target gene activation, but the precise function of and interplay between these factors is not known. This report investigates the Notch IC-interacting proteins, p300, PCAF, and Mastermind-like 1 (MAML1), in an in vitro transcription system with purified factors and naked DNA or chromatin templates. MAML1, RBP-J{kappa}, and Notch IC are all required for optimal transcription from DNA, whereas transcription from chromatin requires, in addition, p300, which interacts with MAML1. The transcriptional activity of p300 requires acetyl coenzyme A, indicating that it functions as a histone acetyltransferase when mediating Notch IC function. PCAF is unable to promote transcription on its own but enhances Notch IC-mediated transcription from chromatin in conjunction with p300. These data define a critical role for p300 in the potentiation of Notch IC function by MAML1 and PCAF, provide the first evidence for cooperativity between PCAF and p300 in Notch IC function, and also indicate direct effects of RBP-J{kappa}, Notch IC, and MAML1 on the general transcription machinery.

* Corresponding author. Mailing address: Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-7604. Fax: (212) 327-7949. E-mail: wallbea{at}mail.rockefeller.edu.

Molecular and Cellular Biology, November 2002, p. 7812-7819, Vol. 22, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.22.7812-7819.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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