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

Minami, Takashi; Murakami, Takeshi; Horiuchi, Keiko; Miura, Mai; Noguchi, T.; Miyazaki, Jun‐ichi; Hamakubo, Takao; Aird, William C.; Kodama, Tatsuhiko

doi:10.1074/jbc.m308730200

Cited by 51 publications

(58 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Western Blot Analysis-Endothelial cells were washed with ice-cold PBS, collected with a cell scraper, and lysed with RIPA buffer as described previously (30). The membrane was blocked with TBS-T containing 2% skim milk and incubated with primary antibodies against GATA3 (Santa Cruz Biotechnology), Tie2 (Santa Cruz Biotechnology), or ␤-actin (Sigma).…”

Section: Methodsmentioning

confidence: 99%

“…The gel was transferred to the membrane, which was incubated either with anti-FLAG antibody or anti-GATA3 antibody (R & D Systems). Alternatively, confluent HUVEC (2.5 ϫ 10 7 cells) were harvested for nuclear extract according to the nitrogen cavitation method to preserve protein-protein associations (30,32). Total nuclear extract was incubated with 30 g of anti-ELF1 antibody (Santa Cruz Biotechnology) or rabbit control IgG (Santa Cruz Biotechnology) overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…For co-immunoprecipitation assays, COS-7 cells were co-transfected with pFLAG-ELF1 and pcDNA3-GATA3 expression plasmids using the FuGENE-HD reagent (Roche Applied Science) as instructed by the manufacturer. Two days later, the transfected cells were lysed, and extracts were prepared as described previously (30). The extracts were incubated with anti-FLAG antibody (Sigma) overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…Plasmids, Transient Transfections, and Luciferase AssaysThe construction of Tie2-luc, Tie2 (GATA mut)-luc, Tie2 (Ets mut)-luc, and expression plasmids for human GATA3 and ELF1 was described previously (25,26,30). HUVEC and HEK-293 cells were transiently transfected with plasmid DNA using FuGENE HD reagent (Roche Applied Science), and luciferase activities were calculated using the Dual-Luciferase assay kit (Promega, Madison, MI) as described previously (31).…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

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

Song

Suehiro

Kanki

et al. 2009

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Endothelial phenotypes are highly regulated in space and time by both transcriptional and post-transcriptional mechanisms. There is increasing evidence that the GATA family of transcription factors function as signal transducers, coupling changes in the extracellular environment to changes in downstream target gene expression. Here we show that human primary endothelial cells derived from large blood vessels express GATA2, -3, and -6. Of these factors, GATA3 was expressed at the highest levels. In DNA microarrays of human umbilical vein endothelial cells (HUVEC), small interfering RNA-mediated knockdown of GATA3 resulted in reduced expression of genes associated with angiogenesis, including Tie2. At a functional level, GATA3 knockdown inhibited angiopoietin (Ang)-1-mediated but not vascular endothelial cell growth factor (VEGF)-mediated AKT signaling, cell migration, survival, and tube formation. In electrophoretic gel mobility shift assays and chromatin immunoprecipitation, GATA3 was shown to bind to regulatory regions within the 5-untranslated region of the Tie2 gene. In co-immunoprecipitation and co-transfection assays, GATA3 and the Ets transcription factor, ELF1, physically interacted and synergized to transactivate the Tie2 promoter. GATA3 knockdown blocked the ability of Ang-1 to attenuate vascular endothelial cell growth factor stimulation of vascular cell adhesion molecule-1 expression and monocytic cell adhesion. Moreover, exposure of human umbilical vein endothelial cells to tumor necrosis factor-␣ resulted in marked down-regulation of GATA3 expression and reduction in Tie2 expression. Together, these findings suggest that GATA3 is indispensable for Ang-1-Tie2-mediated signaling in large vessel endothelial cells.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

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

Song

Suehiro

Kanki

et al. 2009

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…GATA-1, GATA-2, and GATA-3 expression constructs were prepared as described in ref. 36. KRR expression construct, a GATA-3 dominantnegative mutant, was a gift from Aster Winoto (27), and ROG expression vector was from Tang K. Tang (37).…”

Section: Methodsmentioning

confidence: 99%

Interaction of GATA-3/T-bet transcription factors regulates expression of sialyl Lewis X homing receptors on Th1/Th2 lymphocytes

Chen

Osada

Santamaria‐Babí

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Selectin-dependent cell adhesion mediates inflammatory extravasation and routine homing of lymphocytes. Most resting peripheral T lymphocytes lack expression of sialyl Lewis X, the carbohydrate ligand for selectins, and are induced to strongly express it upon activation. T helper 1 (Th1) cells are known to more preferentially express sialyl Lewis X as compared with T helper 2 (Th2) cells upon activation. The molecular basis for this preferential expression, however, has not been elucidated to date. Here we show that the gene for fucosyltransferase VII (FUT7), the rate-limiting enzyme for sialyl Lewis X synthesis, is a unique example of the human genes with binding sites for both GATA-3 and T-bet, two opposing factors for Th1 and Th2 development, and is regulated transcriptionally by a balance of the two interacting transcription factors. T-bet promotes and GATA-3 represses FUT7 transcription. Our results indicated that T-bet interferes with the binding of GATA-3 to its target DNA, and also that GATA-3 significantly interferes with the binding of T-bet to the FUT7 promoter. T-bet has a binding ability to GATA-3, CBP͞P300, and Sp1 to form a transcription factor complex, and GATA-3 regulates FUT7 transcription by phosphorylationdependently recruiting histone deacetylase (HDAC)-3͞HDAC-5 and by competing with CBP͞P300 in binding to the N terminus of T-bet. Suppression of GATA-3 activity by dominant-negative GATA-3 or repressor of GATA (ROG) was necessary to attain a maximum expression of FUT7 and sialyl Lewis X in human T lymphoid cells. These results indicate that the GATA-3͞T-bet transcription factor complex regulates the cell-lineage-specific expression of the lymphocyte homing receptors.fucosyltransferase ͉ helper memory T cells ͉ lymphocyte homing ͉ selectin ͉ GATA-3͞HDAC-3 interaction

show abstract

Colorectal carcinoma cell production of transforming growth factor beta decreases expression of endothelial cell vascular endothelial growth factor receptor 2

Kuczynski

MSc

Coomber

2011

Cancer

View full text Add to dashboard Cite

BACKGROUND: Vascular endothelial growth factor (VEGF) signaling is a target for antiangiogenic cancer therapy. The authors have previously observed that up to 40% of vessels in colorectal carcinoma (CRC) tumors are negative for VEGF receptor 2 (VEGFR2) expression. Differential activity of transforming growth factor beta (TGF-b) is a potential contributor to this receptor heterogeneity because TGF-b contributes to both angiogenesis and CRC tumor progression. METHODS: The authors analyzed VEGFR2 expression by Western blotting, and TGF-b expression in endothelial and CRC cell lines, respectively. In addition, they immunostained endothelial cells in CRC xenografts to find an association between VEGFR2 and TGF-b levels or activity. RESULTS: In bovine aortic endothelial cells (BAECs), TGF-b1 significantly repressed VEGFR2 protein in a time-dependent and dose-dependent fashion (P < .05). Serum-free conditioned media from various malignant human CRC cell lines (HCT116, 379.2, Dks8, and DLD1) induced down-regulation of VEGFR2 in BAECs. This effect was proportional to the total levels of TGF-b1 and TGF-b2 and was blocked by SB-431542 and SD-208, TGF-b receptor I inhibitors. Immunofluorescence staining of subcutaneous mouse xenografts of HCT116, 379.2, Dks8, and SW480 cells revealed vessels with an inverse relationship between TGF-b activity and VEGFR2 expression. Oxygen and bone morphogenetic protein 9 levels were shown to modulate TGF-binduced VEGFR2 down-regulation. CONCLUSIONS: In combination with other factors, TGF-b may contribute to the vascular heterogeneity in human colorectal tumors.

show abstract

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

Cited by 51 publications

References 48 publications

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

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

Interaction of GATA-3/T-bet transcription factors regulates expression of sialyl Lewis X homing receptors on Th1/Th2 lymphocytes

Colorectal carcinoma cell production of transforming growth factor beta decreases expression of endothelial cell vascular endothelial growth factor receptor 2

Contact Info

Product

Resources

About