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J. Biol. Chem., Vol. 279, Issue 7, 5460-5469, February 13, 2004

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Regulation of Immunoglobulin Promoter Activity by TFII-I Class Transcription Factors^*

Dean Tantin, Maria Isabel Tussie-Luna¶, Ananda L. Roy¶, and Phillip A. Sharp||**

From the Department of Biology and Center for Cancer Research, Massachusetts Institute of Technology, ^||McGovern Institute, Cambridge, Massachusetts 02139-4307, and the ^¶Department of Pathology, Tufts University School of Medicine, Boston, Massachusetts 02111

The restriction of immunoglobulin variable region promoter activity to B lymphocytes is a well known paradigm of promoter specificity. Recently, a cis-element, located downstream of the transcription initiation site of murine heavy chain variable promoters, was shown to be critical for B cell activity and specificity. Here we show that mutation of this element, termed DICE (Downstream Immunoglobulin Control Element), reduces in vivo activity in B cells. Gel mobility shift assays show that DICE forms B cell-specific complexes that were also sensitive to DICE mutation. DICE mutation strongly reduces the ability of a distal immunoglobulin heavy chain intronic enhancer to stimulate transcription. We also identify a DICE-interacting factor: a TFII-I-related protein known as BEN (also termed Mus-TRD1 and WBSCR11). Dominant-negative and RNA_i-mediated knockdown experiments indicate that BEN can both positively and negatively regulate IgH promoter activity, depending on the cell line.

Received for publication, October 10, 2003

^* This work was supported in part by United States Public Health Service Grants AI 45150 (to A. L. R.) and PO1-CA42063 (to P. A. S.) and Cancer Center Support (core) Grant P30-CA14051 from the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by fellowships from the Irvington Institute for Immunological Research and The Medical Foundation, Charles A. King Trust.

^** To whom correspondence should be addressed: Dept. of Biology and Center for Cancer Research, Massachusetts Institute of Technology, McGovern Institute, 77 Massachusetts Ave., Rm. E17-529, Cambridge, MA 02139-4307. Tel.: 617-253-6421; Fax: 617-253-3867; E-mail: sharppa{at}mit.edu.

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