Function of GATA Transcription Factors in Induction of Endothelial Vascular Cell Adhesion Molecule-1 by Tumor Necrosis Factor-α

Abstract

Abstract-Endothelial vascular cell adhesion molecule-1 (VCAM-1) is expressed in response to cytokine stimulation and plays a critical role in inflammatory reactions. Previously, we developed a novel VCAM-1 inhibitor that acts through a mechanism independent of nuclear factor-B activity. It suppresses the binding activity of GATA proteins in cytokine-stimulated endothelial cells, which may be related to the anti-VCAM-1 induction effect of this drug. In this study, we investigated the role of GATA proteins in the induction of VCAM-1 by tumor necrosis factor-␣ (TNF-␣) in human endothelial cells. The mRNA expression of GATA-6 was increased, whereas GATA-3 mRNA was decreased by TNF-␣ stimulation. Electrophoretic mobility shift assay showed that TNF-␣ stimulation increased the DNA binding of GATA-6 but decreased that of GATA-3. Experiments using protein overexpression or antisense oligonucleotides revealed that GATA-6 potently acts as a positive regulator of VCAM-1 gene transcription. In contrast, overexpression of GATA-3 was able to suppress TNF-␣-induced VCAM-1 expression. Our results provide evidence of the importance of GATA proteins in the induction of VCAM-1 by TNF-␣ in vascular endothelial cells. The switch from GATA-3 to GATA-6 is taken to be an important transcriptional control event in TNF-␣ induction of VCAM-1. Key Words: endothelial cells Ⅲ vascular cell adhesion molecule-1 Ⅲ GATA transcription factor Ⅲ tumor necrosis factor-␣ Ⅲ human umbilical vein endothelial cells A dherence to the blood vessel wall via cell adhesion molecules is an important step in leukocyte migration. 1 Vascular cell adhesion molecule-1 (VCAM-1) is a cell surface glycoprotein that belongs to the immunoglobulin superfamily, and its expression is dependent on cytokine stimulation. In the vascular system, VCAM-1 is expressed on activated endothelial cells, smooth muscle cells, and fibroblasts in a variety of pathological conditions, including atherosclerosis. [2][3][4] In the VCAM-1 gene regulatory region, functional transcription factor binding motifs have been reported, including nuclear factor (NF)-B, stimulatory protein-1, interferon regulatory factor, activator protein-1, and GATA. 5-8 GATA proteins are members of a zinc finger protein family of transcription factors that recognize the consensus (A/T) GATA(A/G) and related sequences. 9 They are essential for the differentiation and function of hematopoietic cells and endothelial cells as well as fetal differentiation of the cardiovascular system. 10 -12 Although several lines of evidence have provided support for the importance of the GATA element in the upstream promoter region of the VCAM-1 gene, 5,8,13,14 the specific regulatory effects exerted by the GATA factors on endothelial gene expression have yet to be elucidated. Recently, we developed a novel VCAM-1 inhibitor, K-7174, which acts through a mechanism independent of NF-B activity. 15 K-7174 suppressed the binding activity of GATA proteins in cytokine-stimulated human umbilical vein endothelial cells (HUVECs), which may be...

Critical Role for GATA3 in Mediating Tie2 Expression and Function in Large Vessel Endothelial Cells

“…Previous studies have demonstrated that HUVEC express GATA2, GATA3, and GATA6 (41). In addition, GATA2 and GATA3 were identified in human coronary artery endothelial cells (42).…”

Interaction between Hex and GATA Transcription Factors in Vascular Endothelial Cells Inhibits flk-1/KDR-mediated Vascular Endothelial Growth Factor Signaling

“…GATA-2, -3, and -6 have highly conserved zinc finger domain and the region involved the usual protein-protein interactions (2,10,13,15,18,19,20,29,37,39). Further studies will be required to determine whether these motifs are responsible for mediating the binding of Hex to GATA-2, GATA-3, and GATA-6.…”

“…The loaded gel was fixed with 10% methanol and 10% acetic acid and then imaged by assayed for luciferase activity. *, p Ͻ 0.002 compared with the activity from co-transfection with KDR-luc, pMT 2 -GATA2, and pcDNA3; §, p ϭ 0.002 compared from co-transfection with KDR (SP1 mut)-luc, pMT 2 , and pcDNA3; ‡, p ϭ 0.08 (not significant) compared from co-transfection with KDR (GATA mut)-luc, pMT 2 , and pcDNA3. The expression levels were normalized to pRL-SV40 activity and expressed as fold induction relative to co-transfection with vector alone.…”

Efficient differentiation of hESCs into endothelial cells in vitro is secured by epigenetic changes

“…The GATA transcription factor family comprises six members and three of them (GATA 1, 2, 3) contribute to different aspects of hematopoietic development. 9 GATA-2 is expressed in different tissues and plays a major role in the differentiation of hematopoietic cell precursors. 9 In the process of differentiation GATA-2 activates eNOS, endothelin-1, Flk-1, PECAM-1, vWF, and other endothelial genes.…”

“…9 GATA-2 is expressed in different tissues and plays a major role in the differentiation of hematopoietic cell precursors. 9 In the process of differentiation GATA-2 activates eNOS, endothelin-1, Flk-1, PECAM-1, vWF, and other endothelial genes. [9][10][11][12][13] The role of GATA-3 in endothelium formation is poorly understood although it does regulate expression of VCAM-1.…”