Core Element Cloning, Cis - Element Mapping and Serum Regulation of the Human EphB4 Promoter: A Novel TATA-Less Inr/MTE/DPE - Like Regulated Gene.

Autor:	Scalia P; Sbarro Institute for Cancer Research and Molecular Medicine, Biology Department, CST, Temple University, Philadelphia, PA 19122, USA.; Istituto Somatogene per la Oncologia Personalizzata e la Ricerca Onco-Genomica, 93100 Caltanissetta, Italy., Williams SJ; Sbarro Institute for Cancer Research and Molecular Medicine, Biology Department, CST, Temple University, Philadelphia, PA 19122, USA.; Somatolink Foundation, Inc, Philadelphia, PA 19102, USA., Giordano A; Sbarro Institute for Cancer Research and Molecular Medicine, Biology Department, CST, Temple University, Philadelphia, PA 19122, USA.; Dept of Medical Biotechnology, University of Siena, 53100 Siena, Italy.
Jazyk:	angličtina
Zdroj:	Genes [Genes (Basel)] 2019 Dec 02; Vol. 10 (12). Date of Electronic Publication: 2019 Dec 02.
DOI:	10.3390/genes10120997
Abstrakt:	The EphB4 gene encodes for a transmembrane tyrosine kinase receptor involved in embryonic blood vessel differentiation and cancer development. Although EphB4 is known to be regulated at the post-translational level, little is known about its gene regulation. The present study describes the core promoter elements' identification and cloning, the cis-regulatory elements' mapping and the serum regulation of the human EphB4 gene promoter region. Using bioinformatic analysis, Sanger sequencing and recombinant DNA technology, we analyzed the EphB4 gene upstream region spanning +40/-1509 from the actual transcription start site (TSS) and proved it to be a TATA-less gene promoter with dispersed regulatory elements characterized by a novel motif-of-ten element (MTE) at positions +18/+28, and a DPE-like motif and a DPE-like-repeated motif (DRM) spanning nt +27/+30 and +32 +35, respectively. We also mapped both proximal (multiple Sp1) and distal (HoxA9) trans-activating/dispersed cis-acting transcription factor (TF)-binding elements on the region we studied and used a transient transfection reporter assay to characterize its regulation by serum and IGF-II using EphB4 promoter deletion constructs with or without the identified new DNA-binding elements. Altogether, these findings shed new light on the human EphB4 promoter structure and regulation, suggesting mechanistic features conserved among Pol-II TATA-less genes phylogenetically shared from Drosophila to Human genomes. Competing Interests: The authors declare no conflict of interest.
Databáze:	MEDLINE
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