Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene
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...
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Veröffentlicht in: | The Journal of biological chemistry 1996-10, Vol.271 (42), p.26320-26328 |
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Zusammenfassung: | 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 5â²-regulatory region and 85 nucleotides of coding sequence; a ~15-kb intron;
and Exon II, which contains the remainder of the coding sequence and the entire 3â²-untranslated region. Multiple transcription
initiation sites were identified by S1 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
~3.2 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. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.271.42.26320 |