Cloning and identification of regulatory sequences of the human thrombin receptor gene

Fengzhi Li, Demir Baykal, Chris Horaist, Chang Ning Yan, Boyd N. Carr, Rao Gadiparthi, Marschall S. Runge

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing. To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene. The HTR gene consists of Exon I, which contains the 5'-regulatory region and 85 nucleotides of coding sequence; a κ15-kb intron; and Exon II, which contains the remainder of the coding sequence and the entire 3'-untranslated region. Multiple transcription initiation sites were identified by SI mapping and ribonuclease protection assay. DNA sequence analysis indicated the presence of two SP- 1-AP-2 consensus binding sequences, near or within the transcription initiation sites, and consensus binding sequences for numerous regulatory proteins that potentially modulate HTR expression. Functional analysis of the HTR promoter was performed by transfecting human microvascular endothelial cells with HTR promoter region-luciferase constructs. The highest level of expression was obtained with a 0.7-kb promoter sequence and was progressively less with fragments of 0.54, 1.16, 1.6, and κ3.2 kb. The data presented in this report provide a foundation for further characterization of the HTR gene and the mechanisms that regulate its expression within the blood vessel wall.

Original languageEnglish (US)
Pages (from-to)26320-26328
Number of pages9
JournalJournal of Biological Chemistry
Volume271
Issue number42
DOIs
StatePublished - Oct 29 1996
Externally publishedYes

Fingerprint

Thrombin Receptors
Cloning
Organism Cloning
Genes
Transcription Initiation Site
Consensus Sequence
Blood Vessels
Exons
Functional analysis
Nucleic Acid Regulatory Sequences
DNA sequences
Endothelial cells
Transcription Factor AP-1
Blood vessels
3' Untranslated Regions
Ribonucleases
Luciferases
DNA Sequence Analysis
Vascular Smooth Muscle
Genetic Promoter Regions

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cloning and identification of regulatory sequences of the human thrombin receptor gene. / Li, Fengzhi; Baykal, Demir; Horaist, Chris; Yan, Chang Ning; Carr, Boyd N.; Gadiparthi, Rao; Runge, Marschall S.

In: Journal of Biological Chemistry, Vol. 271, No. 42, 29.10.1996, p. 26320-26328.

Research output: Contribution to journalArticle

Li, Fengzhi ; Baykal, Demir ; Horaist, Chris ; Yan, Chang Ning ; Carr, Boyd N. ; Gadiparthi, Rao ; Runge, Marschall S. / Cloning and identification of regulatory sequences of the human thrombin receptor gene. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 42. pp. 26320-26328.
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AU - Gadiparthi, Rao

AU - Runge, Marschall S.

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