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J. Biol. Chem., Vol. 279, Issue 43, 45028-45035, October 22, 2004

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Differential Functions for the Transcription Factor E2A in Positive and Negative Gene Regulation in Pre-B Lymphocytes^*

Stephen Greenbaum, Adam S. Lazorchak, and Yuan Zhuang

From the Department of Immunology, Duke University Medical Center, Durham, North Carolina 27710

The transcription factors encoded by the E2A gene have been shown to play essential roles in the initiation and progression of lymphocyte development. However, there is still a lack of comprehensive understanding of E2A downstream genes in B-cell development. We previously developed a gene tagging-based chromatin immunoprecipitation (ChIP) system to directly evaluate E2A target genes in B-cell development. Here, we have improved this ChIP strategy and used it in conjunction with microarray analysis on E2A-deficient pre-B-cell lines to determine E2A target genes in lymphocyte development. Both microarray data and ChIP studies confirmed that E2A directly controls IgH gene expression. The microarray assay also revealed genes that were significantly up-regulated after E2A disruption. ChIP analysis showed that E2A was most likely to be directly involved in repression of some of these target genes such as Nfil3 and FGFR2. An inducible E2A reconstitution system further demonstrated that E2A-mediated repression of Nfil3 and FGFR2 was reversible. Collectively, these findings indicate that E2A is a positive regulator for one set of genes and a negative regulator for another set of genes in developing B lymphocytes.

Received for publication, January 5, 2004 , and in revised form, August 12, 2004.

^* This work was supported by research Grant R01 CA72433 from the National Institute of Health and a scholarship from the Leukemia and Lymphoma Society Ref. 1485-99 (to Y. Z.). 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.

These authors contributed equally to this work.

To whom correspondence should be addressed: Dept. of Immunology, Duke University Medical Center, Box 3010, 328 Jones Bldg., Research Dr., Durham, NC 27710. Tel.: 919-613-7824; E-mail: yzhuang{at}acpub.duke.edu.

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