Cloning of the rat tissue factor cDNA and promoter: identification of a serum-response region.
Tissue factor (TF) initiates coagulation and its expression in vascular smooth muscle cells (VSMC) likely plays a role in the propagation of arterial thrombosis. We report cloning the cDNA and proximal promoter region of the rat TF gene. While maintaining the general structure and organization of the TF molecule, there is a surprising divergence (approximately 18%) between the derived amino acid sequences of the rat and mouse TF. In contrast, there is striking similarity (90%) in the 5' untranslated regions. High levels of basal promoter activity were seen in rat VSMC with constructs containing 106 bp of sequence downstream from the putative transcription start site and 426 to 103 bp of upstream sequence. Deletion of the sequence from -103 to -79, containing a single SP1 site, removed virtually all of the basal and serum-induced activity. Removal of the NF kappa B site or two additional upstream SP1 sites had little effect on serum responsiveness. Removal of the 5' untranslated region abolished most of the basal activity of the TF promoter, suggesting that its high degree of conservation may be due to the presence of transcriptional elements critical for TF expression in rodent VSMC.
Taby
O
O
Department of Medicine (Divisions of Cardiology and Thrombosis Research), Mount Sinai School of Medicine, New York, NY 10029, USA.
Rosenfield
C L
CL
Bogdanov
V
V
Nemerson
Y
Y
Taubman
M B
MB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051381:Rats / Q000235:genetics*; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D017384:Sequence Deletion; D017385:Sequence Homology; D013045:Species Specificity; D013925:Thromboplastin / Q000235:genetics*
D018076:DNA, Complementary; D013925:Thromboplastin