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Molecular and Cellular Biology, July 2001, p. 4337-4346, Vol. 21, No. 13
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.13.4337-4346.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

A Human Protein with Sequence Similarity to Drosophila Mastermind Coordinates the Nuclear Form of Notch and a CSL Protein To Build a Transcriptional Activator Complex on Target Promoters

Motoo Kitagawa,1,* Toshinao Oyama,1,2 Taichi Kawashima,1,dagger Barry Yedvobnick,3 Anumeha Kumar,3 Kenji Matsuno,4 and Kenichi Harigaya1

Department of Molecular and Tumor Pathology, Chiba University Graduate School of Medicine, Chuo-ku, Chiba 260-8670,1 Graduate School of Science and Technology, Chiba University, Inage-ku, Chiba 263-8522,2 and Department of Biological Science and Technology, Science University of Tokyo, Noda, Chiba 278-8510,4 Japan, and Department of Biology, O. Wayne Rollins Research Center, Emory University, Atlanta, Georgia 303223

Received 29 January 2001/Returned for modification 16 March 2001/Accepted 3 April 2001

Mastermind (Mam) has been implicated as an important positive regulator of the Notch signaling pathway by genetic studies using Drosophila melanogaster. Here we describe a biochemical mechanism of action of Mam within the Notch signaling pathway. Expression of a human sequence related to Drosophila Mam (hMam-1) in mammalian cells augments induction of Hairy Enhancer of split (HES) promoters by Notch signaling. hMam-1 stabilizes and participates in the DNA binding complex of the intracellular domain of human Notch1 and a CSL protein. Truncated versions of hMam-1 that can maintain an association with the complex behave in a dominant negative fashion and depress transactivation. Furthermore, Drosophila Mam forms a similar complex with the intracellular domain of Drosophila Notch and Drosophila CSL protein during activation of Enhancer of split, the Drosophila counterpart of HES. These results indicate that Mam is an essential component of the transcriptional apparatus of Notch signaling.

* Corresponding author. Mailing address: Department of Molecular and Tumor Pathology, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. Phone: 81-43-226-2055. Fax: 81-43-226-2058. E-mail: kitagawa{at}med.m.chiba-u.ac.jp.

dagger Present address: Department of Academic Surgery, Chiba University Graduate School of Medicine, Chuo-ku, Chiba 260-8670, Japan.

Molecular and Cellular Biology, July 2001, p. 4337-4346, Vol. 21, No. 13
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.13.4337-4346.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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