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Molecular and Cellular Biology, March 2009, p. 1163-1175, Vol. 29, No. 5
0270-7306/09/$08.00+0     doi:10.1128/MCB.01572-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Differential Contribution of the Gata1 Gene Hematopoietic Enhancer to Erythroid Differentiation{triangledown} ,{dagger}

Mikiko Suzuki,1,3 Takashi Moriguchi,1 Kinuko Ohneda,2,3 and Masayuki Yamamoto1,3*

Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, and Environmental Response Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Sendai 980-8575,1 Department of Pharmacy, Takasaki University of Health and Welfare, Takasaki 370-0033,2 Center for Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba 305-8577, Japan3

Received 7 October 2008/ Returned for modification 23 November 2008/ Accepted 14 December 2008

GATA1 is a key regulator of erythroid cell differentiation. To examine how Gata1 gene expression is regulated in a stage-specific manner, transgenic mouse lines expressing green fluorescent protein (GFP) reporter from the Gata1 locus in a bacterial artificial chromosome (G1BAC-GFP) were prepared. We found that the GFP reporter expression faithfully recapitulated Gata1 gene expression. Using GFP fluorescence in combination with hematopoietic surface markers, we established a purification protocol for two erythroid progenitor fractions, referred to as burst-forming units-erythroid cell-related erythroid progenitor (BREP) and CFU-erythroid cell-related erythroid progenitor (CREP) fractions. We examined the functions of the Gata1 gene hematopoietic enhancer (G1HE) and the highly conserved GATA box in the enhancer core. Both deletion of the G1HE and substitution mutation of the GATA box caused almost complete loss of GFP expression in the BREP fraction, but the CREP stage expression was suppressed only partially, indicating the critical contribution of the GATA box to the BREP stage expression of Gata1. Consistently, targeted deletion of G1HE from the chromosomal Gata1 locus provoked suppressed expression of the Gata1 gene in the BREP fraction, which led to aberrant accumulation of BREP stage hematopoietic progenitor cells. These results demonstrate the physiological significance of the dynamic regulation of Gata1 gene expression in a differentiation stage-specific manner.

* Corresponding author. Mailing address: Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai 980-8575, Japan. Phone: 81-22-717-8088. Fax: 81-22-717-8090. E-mail: masi{at}mail.tains.tohoku.ac.jp

{triangledown} Published ahead of print on 22 December 2008.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, March 2009, p. 1163-1175, Vol. 29, No. 5
0270-7306/09/$08.00+0     doi:10.1128/MCB.01572-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ohneda, K., Ohmori, S., Ishijima, Y., Nakano, M., Yamamoto, M. (2009). Characterization of a Functional ZBP-89 Binding Site That Mediates Gata1 Gene Expression during Hematopoietic Development. J. Biol. Chem. 284: 30187-30199 [Abstract] [Full Text]  


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