GATA motifs regulate early hematopoietic lineage-specific expression of the Gata2 gene.
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.
Kobayashi-Osaki
Maki
M
Center for Tsukuba Advanced Research Alliance, University of Tsukuba, Japan.
Ohneda
Osamu
O
Suzuki
Norio
N
Minegishi
Naoko
N
Yokomizo
Tomomasa
T
Takahashi
Satoru
S
Lim
Kim-Chew
KC
Engel
James Douglas
JD
Yamamoto
Masayuki
M
D020816:Amino Acid Motifs; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D002460:Cell Line; D019070:Cell Lineage; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology*; D005091:Exons; D005434:Flow Cytometry; D050989:GATA2 Transcription Factor; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D005838:Genotype; D018095:Germ-Line Mutation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D008954:Models, Biological; D008957:Models, Genetic; D009154:Mutation; D011401:Promoter Regions, Genetic; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000737:chemistry* / Q000502:physiology*; D019076:Transgenes
D018952:Antigens, CD34; D017931:DNA Primers; D004268:DNA-Binding Proteins; D050989:GATA2 Transcription Factor; C494711:GATA2 protein, human; C494712:Gata2 protein, mouse; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D004247:DNA; D019009:Proto-Oncogene Proteins c-kit