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Mol Cell Biol, April 1998, p. 2077-2088, Vol. 18, No. 4
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

NF-kappa B2 Is a Putative Target Gene of Activated Notch-1 via RBP-Jkappa

Franz Oswald,1 Susanne Liptay,2 Guido Adler,1 and Roland M. Schmid1,*

Departments of Internal Medicine1 and Pediatrics,2 University of Ulm, D-89081 Ulm, Germany

Received 18 September 1997/Returned for modification 11 November 1997/Accepted 16 January 1998

NF-kappa B2 (p100/p52), a member of the NF-kappa B/Rel family of transcription factors, is involved in the regulation of a variety of genes important for immune function. Previously, we have shown that the NF-kappa B2 gene is regulated in a positive and a negative manner. Two kappa B elements within the NF-kappa B2 promoter mediate tumor necrosis factor alpha-inducible transactivation. In addition, we have shown that there exists a transcriptional repression in the absence of NF-kappa B. To identify a DNA binding activity responsible for this transcriptional repression, we have partially purified a nuclear complex, named Rep-kappa B. Here we further analyze this putative repressive binding activity. Detailed examination of Rep-kappa B-DNA interaction revealed the sequence requirements for binding to be almost identical to those of recombination signal binding protein Jkappa (RBP-Jkappa ), the mammalian homolog of the protein encoded by Drosophila suppressor of hairless [Su(H)]. In addition, in electromobility shift assays, Rep-kappa B binding activity is recognized by an antibody directed against RBP-Jkappa . By performing transient-transfection assays, we show that human RBP-Jkappa represses basal as well as RelA (p65)-stimulated NF-kappa B2 promoter activity. Studies in Drosophila melanogaster have shown that Su(H) is implicated in the Notch signaling pathway regulating cell fate decisions. In transient-transfection assays we show that truncated Notch-1 strongly induces NF-kappa B2 promoter activity. In summary, our data clearly demonstrate that Rep-kappa B is closely related or identical to RBP-Jkappa . RBP-Jkappa is a strong transcriptional repressor of NF-kappa B2. Moreover, this repression can be overcome by activated Notch-1, suggesting that NF-kappa B2 is a novel putative Notch target gene.

* Corresponding author. Mailing address: Department of Internal Medicine, University of Ulm, Robert-Koch-Strasse 8, D-89081 Ulm, Germany. Phone: 49-731-502-4305. Fax: 49-731-502-4302. E-mail: roland.schmid{at}medizin.uni-ulm.de.




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