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Nucleic Acids Research, 1992, Vol. 20, No. 2 287-293
© 1992

MOLECULAR BIOLOGY

Identification of USF as the ubiquitous murine factor that binds to and stimulates transcription from the immunoglobulin {lambda}2 chain promoter

Ling A. Chang, Tammy Smith, Philippe Pognonec1, Robert G. Roeder1 and Helios Murialdo*

Department of Molecular and Medical Genetics, University of Toronto Toronto, Ontario M5S 1A8, Canada 1Laboratory of Biochemistry and Molecular Biology, The Rockefeller University New York, NY 10021, USA

*To whom correspondence should be addressed

Received September 18, 1991. Revised December 16, 1991. Accepted December 16, 1991.

To study the specificity and identity of NF-{lambda}2, a ubiquitous murine nuclear factor that interacts specifically with the promoter of the {lambda}2-chain gene and stimulates its transcription, competition experiments were carried out using DNA fragments from various immunoglobulin regulatory elements. The results showed that a fragment containing the H-chain enhancer competed efficiently for the bonding of NF-{lambda}2. Dissection of the H-chain enhancer revealed that the µE3 motif contributed the competing ability. Additionally, a regulatory region found in the adenovirus major late promoter, which interacts with the human general transcription factor USF, competed very efficiently for bonding to NF-{lambda}2. This region contains a sequence, CACGTGAC, which is identical to a region within the NF-{lambda}2 motif. The pattern of complexes formation using oligonucleotide probes corresponding to the NF-{lambda}2 and USF motifs were identical, and they both differed from that displayed by the E3 probes. Antisera against different domains of USF also react specifically with NF-{lambda}2 showing that this factor is antigenically related, if not identical, to USF. Furthermore, the activity of the {lambda}2 promoter in an in vitro transcription assay was significantly reduced when the nuclear extract used was USF-depleted. Addition of exogenous USF to this extract restored the transcription activity. Therefore, we conclude that NF-{lambda}2 is the murine homologue of USF.


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