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GATA Motifs Regulate Early Hematopoietic Lineage-Specific Expression of the Gata2 Gene

Center for Tsukuba Advanced Research Alliance,¹ JST-ERATO Environmental Response Project,² Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba 305-8577, Japan,³ Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor 48109-06164⁴

Received 1 January 2005/ Returned for modification 3 March 2005/ Accepted 16 May 2005

Transcription factor GATA-2 is essential for definitive hematopoiesis, which developmentally emerges from the para-aortic splanchnopleura (P-Sp). The expression of a green fluorescent protein (GFP) reporter placed under the control of a 3.1-kbp Gata2 gene regulatory domain 5' to the distal first exon (IS) mirrored that of the endogenous Gata2 gene within the P-Sp and yolk sac (YS) blood islands of embryonic day (E) 9.5 murine embryos. The P-Sp- and YS-derived GFP⁺ fraction of flow-sorted cells dissociated from E9.5 transgenic embryos contained far more CD34⁺/c-Kit⁺ cells than the GFP^– fraction did. When cultured in vitro, the P-Sp GFP⁺ cells generated both immature hematopoietic and endothelial cell clusters. Detailed transgenic mouse reporter expression analyses demonstrate that five GATA motifs within the 3.1-kbp Gata2 early hematopoietic regulatory domain (G2-EHRD) were essential for GFP expression within the dorsal aortic wall, where hemangioblasts, the earliest precursors possessing both hematopoietic and vascular developmental potential, are thought to reside. These results thus show that the Gata2 gene IS promoter is regulated by a GATA factor(s) and selectively marks putative hematopoietic/endothelial precursor cells within the P-Sp.

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