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Molecular and Cellular Biology, March 2000, p. 1546-1552, Vol. 20, No. 5
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Recruitment of CREB Binding Protein Is Sufficient for CREB-Mediated Gene Activation

Jean-Rene Cardinaux,1 John C. Notis,1 Qinghong Zhang,1 Ngan Vo,1 Johanna C. Craig,1 Daniel M. Fass,1 Richard G. Brennan,2 and Richard H. Goodman1,*

Vollum Institute1 and Department of Biochemistry,2 Oregon Health Sciences University, Portland, Oregon 97201

Received 14 September 1999/Returned for modification 21 October 1999/Accepted 16 November 1999

Phosphorylation of the transcription factor CREB leads to the recruitment of the coactivator, CREB binding protein (CBP). Recent studies have suggested that CBP recruitment is not sufficient for CREB function, however. We have identified a conserved protein-protein interaction motif within the CBP-binding domains of CREB and another transcription factor, SREBP (sterol-responsive element binding protein). In contrast to CREB, SREBP interacts with CBP in the absence of phosphorylation. We have exploited the conservation of this interaction motif to test whether CBP recruitment to CREB is sufficient for transcriptional activation. Substitution of six nonconserved amino acids from SREBP into the activation domain of CREB confers high-affinity, phosphorylation-independent CBP binding. The mutated CREB molecule, CREBDIEDML, activates transcription in F9 teratocarcinoma and PC12 cells even in the absence of protein kinase A (PKA). Addition of exogenous CBP augments the level of transcription mediated by CREBDIEDML, and adenovirus 12S E1A blocks transcription, implicating CBP in the activation process. Thus, recruitment of CBP to CREB is sufficient for transcriptional activation. Addition of PKA stimulates transcription induced by CREBDIEDML further, suggesting that a phosphorylation event downstream from CBP recruitment augments CREB signaling.

* Corresponding author. Mailing address: Vollum Institute, Oregon Health Sciences University, 3181 S.W. Sam Jackson Park Road, Portland, OR 97201. Phone: (503) 494-5078. Fax: (503) 494-4353. E-mail: goodmanr{at}ohsu.edu.

Molecular and Cellular Biology, March 2000, p. 1546-1552, Vol. 20, No. 5
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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