title: Cloning and structural characterization of the human endothelial nitric-oxide-synthase gene.
abstract: Nitric oxide accounts for the activity of endothelium-derived relaxing factor, which seems to have an important role in vasodilation and inhibition of platelet aggregation. In endothelial cells, one isoform of nitric-oxide synthase is constitutively expressed. Analysis of the cDNA encoding the human endothelial nitric-oxide synthase revealed that the mRNA is 4.1 kb in size and that the translated protein consists of 1203 amino acids. We have cloned a genomic DNA encoding the human endothelial nitric-oxide synthase and analyzed the entire nucleotide sequence of the gene. The gene consists of 26 exons with a total size of 21 kb. The 5' flanking region of the gene lacks TATA boxes, but it contains putative Sp1-binding sites in (G+C)-rich regions. Of particular interest is the fact that a shear-stress-responsive element is located at position -985, which probably regulates the nitric-oxide-synthase gene in response to fluid mechanical forces at the transcriptional level in the vascular endothelium. Two minisatellite sequences are detectable in introns 2 and 8; a 32-bp consensus sequence repeats 38 times and a 57-bp consensus sequence repeats ten times. We found polymorphisms of the BamHI fragment containing the former minisatellite sequence in genomic DNA from pedigree family members. Furthermore, five tandem repeats of a 27-bp core consensus sequence and 35 repeats of a dinucleotide (CA) are located in introns 4 and 13, respectively. These repeat sequences will probably provide genetic markers for gene mapping and linkage analysis of inherited diseases including cardiovascular diseases.
authors.affiliation: Department of Medical Chemistry, Kochi Medical School, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Miyahara
authors.forename: T
authors.initials: T
authors.lastname: Kawamoto
authors.forename: K
authors.initials: K
authors.lastname: Sase
authors.forename: Y
authors.initials: Y
authors.lastname: Yui
authors.forename: K
authors.initials: K
authors.lastname: Toda
authors.forename: L X
authors.initials: LX
authors.lastname: Yang
authors.forename: R
authors.initials: R
authors.lastname: Hattori
authors.forename: T
authors.initials: T
authors.lastname: Aoyama
authors.forename: Y
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Doi
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004276:DNA, Satellite; D004730:Endothelium, Vascular / Q000201:enzymology*; D005091:Exons / Q000235:genetics; D005819:Genetic Markers; D006801:Humans; D007438:Introns / Q000235:genetics; D007527:Isoenzymes / Q000235:genetics*; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
chemical_list: D004276:DNA, Satellite; D005819:Genetic Markers; D007527:Isoenzymes; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases