EDF-1, a Novel Gene Product Down-regulated in Human Endothelial Cell Differentiation

Abstract

Endothelial cell differentiation is a crucial step in angiogenesis. Here we report the identification of EDF-1, a novel gene product that is down-regulated when endothelial cells are induced to differentiate in vitro. The cDNA encoding EDF-1 was isolated by RNA fingerprinting from human endothelial cells exposed to human immunodeficiency virus type 1 Tat, a viral protein known to be angiogenic. The deduced amino acid sequence of EDF-1 encodes a basic intracellular protein of 148 amino acids that is homologous to MBF1 (multiprotein-bridging factor 1) of the silkworm Bombyx mori and to H7, which is implicated in the early developmental events of Dictyostelium discoideum. Interestingly, human immunodeficiency virus type 1 Tat, which affects endothelial functions, and the phorbol ester 12-Otetradecanoylphorbol-13-acetate and culture on fibrin gels, which promote endothelial differentiation in vitro, all down-regulate EDF-1 expression both at the RNA and protein levels. In addition, the inhibition of EDF-1 translation by an antisense anti-EDF-1 construct results in the inhibition of endothelial cell growth and in the transition from a nonpolar cobblestone phenotype to a polar fibroblast-like phenotype. These data suggest that EDF-1 may play a role in the regulation of human endothelial cell differentiation.

Transforming Growth Factor β2 inhibition of Hepatocyte Growth Factor-induced endothelial proliferation and migration

“…To obtain cDNAs representing mRNAs regulated by HGF in human endothelial cells, we utilized a modi®ed PCR-based di erential screening approach commonly referred to as RNA ®ngerprinting (Consalez et al, 1996;Dragoni et al, 1998). We obtained several di erentially expressed clones.…”

“…Several di erentially expressed genes were identi®ed. The bands corresponding to di erentially expressed genes were excised from the polyacrylamide gel, electroeluted, re-ampli®ed by PCR, cloned in Bluescript and sequenced as described (Consalez et al, 1996;Dragoni et al, 1998). Double-stranded sequence analysis was performed using the T7 Sequencing kit (Pharmacia) following the manufacturer's instruction.…”

“…Cell culture, proliferation and migration assays HUVEC-C were from ATCC and cultured in M199 containing 10% FCS, ECGS (150 mg/ml) and heparin (5 U/ ml) (growing medium) on gelatin coated dishes (Dragoni et al, 1998). Bovine aortic endothelial cells (BAE) and bovine adrenal medulla capillary endothelial cells (BCE) (both provided by P Mignatti, New York, USA) were grown in dMEM supplemented with 10% FCS on gelatin-coated dishes.…”

Differential expression and interaction of transcription co-activator MBF1 with TATA-binding protein (TBP) in the apicomplexan Cryptosporidium parvum

“…The CpMBF1 transcription level is low in free sporozoites and intracellular parasites during their early invasion stages, and CpMBF1 protein is barely detectable in intact oocysts and free sporozoites, suggesting that CpMBF1 may play a role in activating certain genes during the intracellular development of C. parvum. In addition to the activation of histidine synthetic genes in yeast under histidine starvation, MBF1-mediated pathways are known to cause cell differentiation in insects, the formation of stalks in the slime mould Dictyostelium discoideum and the differentiation of endothelial cells in mammals (Dragoni et al, 1998;Kabe et al, 1999;Singleton et al, 1991;Takemaru et al, 1997). These observations, together with the parasite life cycle stage-dependent expression pattern, imply that CpMBF1 may be involved in the cell differentiation of C. parvum during its complex life cycle.…”

“…Drosophila nuclear receptor FTZ-F1 or yeast leucine zipper protein GCN4) (Dragoni et al, 1998;Kabe et al, 1999;Li et al, 1994;Liu et al, 2003;Takemaru et al, 1997Takemaru et al, , 1998. We have previously identified its C. parvum counterpart and demonstrated that the CpMBF1 gene is able to functionally complement MBF1 deficiency in yeast (Zhu et al, 2000).…”