GATA motifs regulate early hematopoietic lineage-specific expression of the Gata2 gene
GATA motifs regulate early hematopoietic lineage-specific expression of the Gata2 gene
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DOI：
10.1128/mcb.25.16.7005-7020.2005
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发表时间：
2005-08-01
影响因子：
5.3
通讯作者：
Yamamoto, M
中科院分区：
文献类型：
--
作者：
Kobayashi-Osaki, M;Ohneda, O;Yamamoto, M
转录因子GATA-2对于最终的造血至关重要，这种造血是从主动脉旁胸膜（P-Sp）发育而来的。在3.1 kbp的Gata2基因调控域5′至远端第一外显子（IS）的控制下，绿色荧光蛋白（GFP）报告基因的表达与胚胎日(E) 9.5小鼠胚胎P-Sp和卵黄囊（YS）血岛内的内源性Gata2基因的表达一致。从E9.5转基因胚胎分离的流动分选细胞中，P-Sp-和ys -衍生的GFP（+）部分比GFP（-）部分含有更多的CD34(+)/c-Kit（+）细胞。在体外培养时，P-Sp GFP（+）细胞产生了未成熟的造血细胞和内皮细胞簇。详细的转基因小鼠报告基因表达分析表明，3.1 kbp GATA a2早期造血调节结构域（G2-EHRD）内的5个GATA基序对于主动脉背壁内GFP的表达至关重要，而主动脉背壁被认为是血管母细胞（具有造血和血管发育潜能的最早的前体）的所在地。这些结果表明，Gata2基因启动子受GATA因子调控，并选择性地标记P-Sp内的推定造血/内皮前体细胞。
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.