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 Blood, 15 September 2005, Vol. 106, No. 6, pp. 1948-1955.
Prepublished online as a Blood First Edition Paper on May 31, 2005; DOI 10.1182/blood-2004-12-4872.

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HEMATOPOIESIS

Involvement of Runx1 in the down-regulation of fetal liver kinase-1 expression during transition of endothelial cells to hematopoietic cells

Hideyo Hirai, Igor M Samokhvalov, Tetsuhiro Fujimoto, Satomi Nishikawa, Jiro Imanishi, and Shin-Ichi Nishikawa

From the Department of Microbiology, Kyoto Prefectural University of Medicine, Kyoto, Japan; RIKEN Center for Developmental Biology, Minatoshimaminamimachi, Kobe, Japan; and European Molecular Biology Laboratory, Monterotondo, Italy.

During early mouse embryogenesis, fetal liver kinase-1 (Flk-1), a receptor for vascular endothelial growth factor, and Runx1, a runt domain transcription factor, have prerequisite roles in the generation of hematopoietic lineages. Flk-1 expression is maintained in successive stages from mesodermal to endothelial cells and is down-regulated in nascent hematopoietic cells, whereas Runx1 (Runt-related transcription factor 1) is expressed in embryonic sites of hematopoietic cell de novo generation and in practically all hematopoietic organs. Here we show that Runx1 represses Flk-1 during the development of hemogenic endothelial cells into hematopoietic cells. We established embryonic stem cell clones carrying the Venus gene, a modified version of yellow fluorescence protein, in the Runx1 locus and cultured them on OP9 cells. Flk-1+ cells appeared on day 3.5, and Runx1+ cells first appeared from the Flk-1+ fraction on day 4.5. The Flk-1+Runx1+ cells rapidly stopped expressing Flk-1 with further incubation and eventually gave rise to CD45+ or TER119+ cells. Runx1 repressed Flk-1 promoter transcriptional activity in an endothelial cell line, and this repression required intact DNA-binding and transactivating domains of Runx1 protein. The repressor activity of Runx1 endogenous Flk-1 was also confirmed overexpressing Runx1 in embryonic stem cell differentiation cultures. These results provide novel insight into the role Runx1 during the development of hematopoietic cell lineages.


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