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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-
B2 Is a Putative Target Gene of Activated Notch-1 via
RBP-J
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-
B2 (p100/p52), a member of the NF-
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-
B2 gene is regulated in a positive and a negative manner. Two
B elements within the NF-
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-
B. To
identify a DNA binding activity responsible for this transcriptional
repression, we have partially purified a nuclear complex, named
Rep-
B. Here we further analyze this putative repressive binding
activity. Detailed examination of Rep-
B-DNA interaction revealed
the sequence requirements for binding to be almost identical to those
of recombination signal binding protein J
(RBP-J
), the mammalian
homolog of the protein encoded by Drosophila suppressor of
hairless [Su(H)]. In addition, in electromobility shift assays,
Rep-
B binding activity is recognized by an antibody directed against
RBP-J
. By performing transient-transfection assays, we show that
human RBP-J
represses basal as well as RelA (p65)-stimulated
NF-
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-
B2 promoter activity. In summary, our data clearly
demonstrate that Rep-
B is closely related or identical to RBP-J
.
RBP-J
is a strong transcriptional repressor of NF-
B2. Moreover,
this repression can be overcome by activated Notch-1, suggesting that
NF-
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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