Alternative Promoters Regulate Transcription of the Mouse GATA-2 Gene

doi:10.1074/jbc.273.6.3625

Alternative Promoters Regulate Transcription of the Mouse GATA-2 Gene*

From the ^‡Center for Tsukuba Advanced Research Alliance and Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba 305 and the ^§Department of Biochemistry, Tohoku University School of Medicine, Seiryocho, Aoba-ku, Sendai 980-77, Japan

Abstract

Transcription factor GATA-2 has been shown to be a key regulator in hematopoietic progenitor cells. To elucidate how the expression of the GATA-2 gene is controlled, we isolated themouse GATA-2 (mGATA-2) gene. Transcription of mGATA-2 mRNAs was found to initiate from two distinct first exons, both of which encode entirely untranslated regions, while the remaining five exons are shared by each of the two divergent mRNAs. Reverse transcriptase-polymerase chain reaction analysis revealed that GATA-2 mRNA initiated at the upstream first exon (IS) in Sca-1⁺/c-kit⁺ hematopoietic progenitor cells, whereas mRNA that initiates at the downstream first exon (IG) is expressed in all tissues and cell lines that express GATA-2. While the structure of the IG exon/promoter shows high similarity to those of the Xenopus and human GATA-2 genes, the IS exon/promoter has not been described previously. When we examined the regulation contributing to IS transcription using transient transfection assays, we found that sequences lying between −79 and −61 are critical for the cell type-specific activity of the IS promoter. DNase I footprinting experiments and electrophoretic mobility shift assays demonstrated the binding of transcription factors to this region. These data indicate that the proximal 80 base pair region of IS promoter is important for the generation of cell type-specific expression of mGATA-2 from the IS exon.

Footnotes

↵* This work was supported in part by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture and Japanese Society for Promotion of Sciences (RFTF96I00202).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) AB007371 and AB009272.
↵¶ To whom correspondence should be addressed. Tel.: 81-298-53-3111; Fax: 81-298-53-6965; E-mail:masiya{at}igaku.md.tsukuba.ac.jp.
↵1 The abbreviations used are: ES, embryonic stem; IS, upstream first exon; IG, downstream first exon; bp, base pair(s); kbp, kilobase pair(s); FACS, fluorescence-activated cell sorter; RT, reverse transcriptase; PCR, polymerase chain reaction; RACE, rapid amplification of cDNA ends; UTR, untranslated region; EMSA, electrophoretic mobility shift assay.
↵2 N. Suwabe, N. Minegishi, and M. Yamamoto, unpublished data.
↵3 H. Ishihara and M. Yamamoto, unpublished observations.
↵4 Version 1.3© 1995 by Yutaka Akiyama at the World Wide Web sitehttp://www.pdap1.tre.rwcp.or.jp/research/db/TFSEARCH.html.
↵5 J. Ohta, N. Minegishi, and M. Yamamoto, unpublished observation.
- Received August 21, 1997.
- Revision received October 22, 1997.
The American Society for Biochemistry and Molecular Biology, Inc.