Ferdous Anwarul, Caprioli Arianna, Iacovino Michelina, Martin Cindy M, Morris Jesse, Richardson James A, Latif Shuaib, Hammer Robert E, Harvey Richard P, Olson Eric N, Kyba Michael, Garry Daniel J
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75244, USA.
Proc Natl Acad Sci U S A. 2009 Jan 20;106(3):814-9. doi: 10.1073/pnas.0807583106. Epub 2009 Jan 7.
Recent studies support the existence of a common progenitor for the cardiac and endothelial cell lineages, but the underlying transcriptional networks responsible for specification of these cell fates remain unclear. Here we demonstrated that Ets-related protein 71 (Etsrp71), a newly discovered ETS family transcription factor, was a novel downstream target of the homeodomain protein, Nkx2-5. Using genetic mouse models and molecular biological techniques, we demonstrated that Nkx2-5 binds to an evolutionarily conserved Nkx2-5 response element in the Etsrp71 promoter and induces the Etsrp71 gene expression in vitro and in vivo. Etsrp71 was transiently expressed in the endocardium/endothelium of the developing embryo (E7.75-E9.5) and was extinguished during the latter stages of development. Using a gene disruption strategy, we found that Etsrp71 mutant embryos lacked endocardial/endothelial lineages and were nonviable. Moreover, using transgenic technologies and transcriptional and chromatin immunoprecipitation (ChIP) assays, we further established that Tie2 is a direct downstream target of Etsrp71. Collectively, our results uncover a novel functional role for Nkx2-5 and define a transcriptional network that specifies an endocardial/endothelial fate in the developing heart and embryo.
最近的研究支持心脏和内皮细胞谱系存在共同祖细胞,但负责这些细胞命运特化的潜在转录网络仍不清楚。在这里,我们证明了Ets相关蛋白71(Etsrp71),一种新发现的ETS家族转录因子,是同源结构域蛋白Nkx2-5的一个新的下游靶点。利用基因小鼠模型和分子生物学技术,我们证明Nkx2-5与Etsrp71启动子中一个进化保守的Nkx2-5反应元件结合,并在体外和体内诱导Etsrp71基因表达。Etsrp71在发育中的胚胎(E7.75-E9.5)的心内膜/内皮中短暂表达,并在发育后期消失。使用基因破坏策略,我们发现Etsrp71突变胚胎缺乏心内膜/内皮谱系且无法存活。此外,利用转基因技术以及转录和染色质免疫沉淀(ChIP)分析,我们进一步确定Tie2是Etsrp71的直接下游靶点。总的来说,我们的结果揭示了Nkx2-5的一种新的功能作用,并定义了一个在发育中的心脏和胚胎中指定心内膜/内皮命运的转录网络。