Regulation of vascular endothelial growth factor D by orphan receptors hepatocyte nuclear factor-4 alpha and chicken ovalbumin upstream promoter transcription factors 1 and 2.
Vascular endothelial growth factor D has recently been linked to the control of lymphangiogenesis and lymphatic metastasis. The molecular determinants regulating vegf-D gene transcription, however, have not yet been identified. After isolation of 2 kb of 5'-flanking DNA of the human vegf-D gene, we identified a novel, atypical direct repeat (DR) element consisting of a consensus half-site (AGGTCA) at -125/-119 and a degenerated DR half-site (ATGTTA) at -99/-94 as sufficient and necessary for vegf-D transcription. The vegf-D DR element is bound and activated by the orphan receptors hepatocyte nuclear factor 4 alpha (HNF-4 alpha) and chicken ovalbumin upstream promoter transcription factor (COUP-TF)-1/COUP-TF2. Additionally, chromatin immunoprecipitation assays identified transcriptional coactivators cyclic AMP-responsive element binding protein-binding protein and glucocorticoid receptor interacting protein 1 at the vegf-D DR element and functional assays confirmed their stimulatory effect on the vegf-D promoter. Histone deacetylase inhibition by trichostatin A led to accumulation of acetylated histones H3/H4 at the vegf-D promoter, up-regulation of vegf-D mRNA levels, and transactivation of vegf-D promoter reporter gene constructs in cancer cell lines. This study for the first time describes the molecular determinants in cis and trans controlling vegf-D gene transcription and identifies interaction of HNF-4 alpha and COUP-TF1/COUP-TF2 with a proximal, atypical DR element as indispensable for vegf-D transcription. Moreover, our findings suggest that epigenetic control of histone acetylation represents an important determinant of vegf-D gene expression in cancer cells. These results provide novel insights into the molecular machinery controlling vegf-D gene expression and may add to a better understanding of the regulation of lymphangiogenesis in vascular development and cancer.
Schäfer
Georgia
G
Laboratory for Angiogenesis and Tumor Metastasis, Charité University Hospital, Berlin, Germany.
Wissmann
Christoph
C
Hertel
Johannes
J
Lunyak
Victoria
V
Höcker
Michael
M
D000107:Acetylation; D001483:Base Sequence; D051838:COUP Transcription Factor I / Q000378:metabolism / Q000502:physiology*; D051837:COUP Transcription Factor II / Q000378:metabolism / Q000502:physiology*; D050882:CREB-Binding Protein / Q000378:metabolism / Q000502:physiology; D005786:Gene Expression Regulation; D051557:Hepatocyte Nuclear Factor 4 / Q000378:metabolism / Q000502:physiology*; D051548:Histone Acetyltransferases / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D051958:Nuclear Receptor Coactivator 2 / Q000378:metabolism / Q000502:physiology; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D015534:Trans-Activators / Q000378:metabolism / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured; D042643:Vascular Endothelial Growth Factor D / Q000235:genetics*
D051838:COUP Transcription Factor I; D051837:COUP Transcription Factor II; D051557:Hepatocyte Nuclear Factor 4; D006657:Histones; C498001:NCOA2 protein, human; D051958:Nuclear Receptor Coactivator 2; D015534:Trans-Activators; D042643:Vascular Endothelial Growth Factor D; D050882:CREB-Binding Protein; C494363:CREBBP protein, human; D051548:Histone Acetyltransferases