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J Biol Chem, Vol. 275, Issue 13, 9425-9432, March 31, 2000

Eosinophil-specific Regulation of gp91^phox Gene Expression by Transcription Factors GATA-1 and GATA-2^*

Dan Yang, Shoichi Suzuki, Li Jun Hao, Yoshito Fujii, Akira Yamauchi, Masayuki Yamamoto^§, Michio Nakamura^¶, and Atsushi Kumatori

From the  Department of Host-defense Biochemistry, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523 and the ^§ Center of Tsukuba Advanced Research Alliance, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8577, Japan

The glycoprotein gp91^phox is an essential component of the phagocyte NADPH oxidase and is expressed in eosinophils, neutrophils, monocytes, and B-lymphocytes. We previously suggested an eosinophil-specific mechanism of gp91^phox gene expression. To elucidate the mechanism, we performed functional assays on deletion mutants of the gp91^phox promoter in various types of gp91^phox-expressing cells. A 10-base pair (bp) region from bp 105 to 96 of the promoter activated transcription of the gene in eosinophilic cells, but not in neutrophilic, monocytic, or B-lymphocytic cells. A 2-bp mutation introduced into the GATA site spanning bp 101 to 96 (98GATA site) of the fragment abolished its activity. Gel shift assays using a GATA competitor and specific antibodies demonstrated that both GATA-1 and GATA-2 specifically bound to the 98GATA site with similar affinities. Individual transfection of GATA-1 and GATA-2 into Jurkat cells, which have neither endogenous GATA-1 nor GATA-2, activated the 105/+12 construct in a 98GATA site-dependent manner. Combined transfection of GATA-1 and GATA-2 activated the promoter less than transfection of GATA-1 alone. These results suggest that GATA-1 is an activator and that GATA-2 is a relative competitive inhibitor of GATA-1 in the expression of the gp91^phox gene in human eosinophils.

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^* This work was supported by grants-in-aid for encouragement of young scientists and for scientific research from the Ministry of Education, Science, Sports, and Culture of Japan and by grants for Center of Excellence and collaboration research from the Institute of Tropical Medicine.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.

^¶ To whom correspondence should be addressed. Tel.: 81-95-849-7848; Fax: 81-95-849-7805; E-mail: nakamura@net.nagasaki-u.ac.jp.

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Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.

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