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J. Biol. Chem., Vol. 279, Issue 39, 40906-40917, September 24, 2004

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Gfi1:Green Fluorescent Protein Knock-in Mutant Reveals Differential Expression and Autoregulation of the Growth Factor Independence 1 (Gfi1) Gene during Lymphocyte Development^*

Raif Yücel, Christian Kosan, Florian Heyd, and Tarik Möröy

From the Institut für Zellbiologie (Tumorforschung), Universitätsklinikum Essen, Virchowstrasse 173, D-45122 Essen, Germany

The Gfi1 gene encodes a 55-kDa transcriptional repressor protein with important functions in T-cell development, in granulopoiesis, and in the regulation of the innate immune response. To follow expression of the Gfi1 gene during the differentiation of specific immune cells, we have generated a mouse mutant in which the Gfi1 coding region is replaced by the gene for the green fluorescent protein (GFP). We found that Gfi1 gene expression is highest in early B-cell subpopulation and differentially expressed during T-cell development with peak levels at stages where pre-TCR or positive/negative selection takes place. Gfi1 is absent in mature B-cells, whereas in peripheral T-cells Gfi1 gene expression is low but rises significantly upon T-cell receptor triggering and decreases again in T-memory cells. Constitutive expression of an lck promoter-driven Gfi1 transgene led to transcriptional silencing of the Gfi1:GFP allele in T-cells. Because Gfi1 was found to occupy genomic sites of its own promoter in thymocytes and was able to repress its own transcription in vitro we propose that transcription of the Gfi1 gene is regulated through an autoregulatory feedback loop.

Received for publication, January 25, 2004 , and in revised form, July 8, 2004.

^* This work was supported by the Deutsche Forschungsgemeinschaft (Grant Mo435/10-4,10-5), the Fonds der chemischen Industrie, the European Community Framework 5 Program, and the IFORES Program of the University of Essen Medical School. 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.

To whom correspondence should be addressed: Institut für Zellbiologie (Tumorforschung), IFZ, Virchowstrasse 173, D-45122 Essen, Germany. Tel.: 49-201-723-3380; Fax: 49-201-723-5904; E-mail: moeroey{at}uni-essen.de.

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