TY - JOUR
T1 - Organization of the bovine gene encoding the endothelial nitric oxide synthase
AU - Venema, Richard C.
AU - Nishida, Ken'ichi
AU - Alexander, R. Wayne
AU - Harrison, David G.
AU - Murphy, T. J.
N1 - Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1994/8/2
Y1 - 1994/8/2
N2 - The bovine endothelial nitric oxide synthase gene plus 2.9 kilobases of 5′-flanking sequence has been isolated and characterized. The gene spans 20 kilobases and contains 26 exons and 25 introns. Two transcription start sites have been determined by primer extension analysis which are located 170 and 240 base pairs upstream, respectively, from the methionine translational initiation codon. Evidence supporting the upstream boundary region for transcriptional initiation was also obtained by reverse transcription-polymerase chain reaction. The 5′-flanking region lacks a typical TATA box but contains numerous putative transcription factor binding sites. These include consensus sequences for an AP-1 site, an NF-1 site, a tumor necrosis factor responsive element, two sterol regulatory elements, 3 acute-phase response elements, 6 GATA motifs, 16 CACCC boxes, 5 Sp1 sites, 15 estrogen half-palindromic motifs, and 9 fluid shear stress-responsive elements. The isolated gene promoter directs basal transcription of a luciferase reporter gene when transiently transfected into bovine aortic endothelial cells. High sequence homology of the promoter region to the human endothelial nitric oxide synthase gene promoter (75% nucleotide identity in 1.6 kilobases of 5′-flanking sequence) suggests evolutionary conservation of transcriptional regulation. Isolation and characterization of the bovine endothelial nitric oxide synthase gene should facilitate further investigation of mechanisms by which gene expression is regulated.
AB - The bovine endothelial nitric oxide synthase gene plus 2.9 kilobases of 5′-flanking sequence has been isolated and characterized. The gene spans 20 kilobases and contains 26 exons and 25 introns. Two transcription start sites have been determined by primer extension analysis which are located 170 and 240 base pairs upstream, respectively, from the methionine translational initiation codon. Evidence supporting the upstream boundary region for transcriptional initiation was also obtained by reverse transcription-polymerase chain reaction. The 5′-flanking region lacks a typical TATA box but contains numerous putative transcription factor binding sites. These include consensus sequences for an AP-1 site, an NF-1 site, a tumor necrosis factor responsive element, two sterol regulatory elements, 3 acute-phase response elements, 6 GATA motifs, 16 CACCC boxes, 5 Sp1 sites, 15 estrogen half-palindromic motifs, and 9 fluid shear stress-responsive elements. The isolated gene promoter directs basal transcription of a luciferase reporter gene when transiently transfected into bovine aortic endothelial cells. High sequence homology of the promoter region to the human endothelial nitric oxide synthase gene promoter (75% nucleotide identity in 1.6 kilobases of 5′-flanking sequence) suggests evolutionary conservation of transcriptional regulation. Isolation and characterization of the bovine endothelial nitric oxide synthase gene should facilitate further investigation of mechanisms by which gene expression is regulated.
KW - (Bovine aortic endothelial cell)
KW - Gene structure
KW - Nitric oxide synthase
KW - Promoter deletional analysis
KW - Transcriptional regulation
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U2 - 10.1016/0167-4781(94)90195-3
DO - 10.1016/0167-4781(94)90195-3
M3 - Article
C2 - 7519447
AN - SCOPUS:0028068430
SN - 0167-4781
VL - 1218
SP - 413
EP - 420
JO - BBA - Gene Structure and Expression
JF - BBA - Gene Structure and Expression
IS - 3
ER -