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Open AccessCommunication

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

1
Sbarro Institute for Cancer Research and Molecular Medicine, Biology Department, CST, Temple University, Philadelphia, PA 19122, USA
2
Istituto Somatogene per la Oncologia Personalizzata e la Ricerca Onco-Genomica, 93100 Caltanissetta, Italy
3
Somatolink Foundation, Inc, Philadelphia, PA 19102, USA
4
Dept of Medical Biotechnology, University of Siena, 53100 Siena, Italy
*
Author to whom correspondence should be addressed.
Genes 2019, 10(12), 997; https://doi.org/10.3390/genes10120997
Received: 18 October 2019 / Revised: 12 November 2019 / Accepted: 27 November 2019 / Published: 2 December 2019
(This article belongs to the Section Molecular Genetics and Genomics)
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.
Keywords: polymerase-II; TATA(box)-less promoter; motif-of-ten element; downstream promoter element; DPE-like repeated motif; Sp1; HoxA9; insulin-like growth factor-II polymerase-II; TATA(box)-less promoter; motif-of-ten element; downstream promoter element; DPE-like repeated motif; Sp1; HoxA9; insulin-like growth factor-II
MDPI and ACS Style

Scalia, P.; Williams, S.J.; Giordano, A. Core Element Cloning, Cis-Element Mapping and Serum Regulation of the Human EphB4 Promoter: A Novel TATA-Less Inr/MTE/DPE-Like Regulated Gene. Genes 2019, 10, 997.

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