Epigenetically coordinated GATA2 binding is necessary for endothelium-specificendomucinexpression

Abstract

GATA2 is well recognized as a key transcription factor and regulator of cell-type specificity and differentiation. Here, we carried out comparative chromatin immunoprecipitation with comprehensive sequencing (ChIP-seq) to determine genome-wide occupancy of GATA2 in endothelial cells and erythroids, and compared the occupancy to the respective gene expression profile in each cell type. Although GATA2 was commonly expressed in both cell types, different GATA2 bindings and distinct cell-specific gene expressions were observed. By using the ChIP-seq with epigenetic histone modifications and chromatin conformation capture assays; we elucidated the mechanistic regulation of endothelial-specific GATA2-mediated endomucin gene expression, that was regulated by the endothelial-specific chromatin loop with a GATA2-associated distal enhancer and core promoter. Knockdown of endomucin markedly attenuated endothelial cell growth, migration and tube formation. Moreover, abrogation of GATA2 in endothelium demonstrated not only a reduction of endothelial-specific markers, but also induction of mesenchymal transition promoting gene expression. Our findings provide new insights into the correlation of endothelial-expressed GATA2 binding, epigenetic modification, and the determination of endothelial cell specificity.

GATA2 is required for lymphatic vessel valve development and maintenance

“…ChIP-Seq studies performed across a range of cell types demonstrate that GATA2 has distinct sites of chromatin occupancy and transcriptional targets in each cell type in which it acts, suggesting tissue-specific transcriptional interactions (8,24,(44)(45)(46). The zinc fingers of GATA2 serve at least 2 key roles: the C-terminal zinc finger mediates binding to WGATAR motifs (47,48), whereas the N-terminal zinc finger mediates protein-protein interactions with transcriptional cofactors and stabilizes DNA binding (37).…”

Endothelial LSP1 Modulates Extravascular Neutrophil Chemotaxis by Regulating Nonhematopoietic Vascular PECAM-1 Expression

“…5A, targeted siRNA silencing of LSP1 in murine SVEC4-10EE2 endothelial cells significantly blunted PECAM-1, but not ICAM-1, mRNA levels, suggesting that LSP1 transcriptionally regulates PECAM-1 expression in endothelial cells. To elucidate the transcriptional regulation of PECAM-1 by LSP1, we explored the participation of the transcription factor GATA-2 that is expressed in microvascular endothelial cells (42). As shown in Fig.…”

“…PECAM-1 transcription was shown to be regulated by the putative TATA-less promoter region that contains relevant EGR-1 and GATA-2 cisregulatory elements (55,57,58). In addition to regulating PECAM-1 expression, GATA-2 is decisive in endothelial-selective gene expression of endothelial NO synthase, endomucin, and VCAM-1 (42,(58)(59)(60). Recently, endothelial GATA-2 was shown to participate in regulating angiogenesis (61) and maintenance of vascular integrity (62).…”

“…The zinc finger transcription factor GATA-2 was initially shown to be crucial in modulating gene expression in megakaryocytic and erythroid lineages (53). Mounting evidence suggests that GATA-2 plays an important role in nonhematopoietic endothelial cells (54,55), where it is associated with PECAM-1 expression and participates in endothelial cell dedifferentiation (42,56). PECAM-1 transcription was shown to be regulated by the putative TATA-less promoter region that contains relevant EGR-1 and GATA-2 cisregulatory elements (55,57,58).…”

Genome-wide Approaches Reveal Functional Vascular Endothelial Growth Factor (VEGF)-inducible Nuclear Factor of Activated T Cells (NFAT) c1 Binding to Angiogenesis-related Genes in the Endothelium

“…2D). These NFATc1 occupancy patterns were unique, considering our previous reports of GATA2-and STAT6-mediated ChIP-seqs in endothelial cells (20,23). GATA2 and STAT6 predominantly occupied distal enhancer regions.…”