Human Tissue Factor: DNA Sequence and Chromosome Localization of the Gene

Eleonora M. Scarpati, Robert R. Flandermeyer, Ned R. Siegel, Duanzhi Wen, J. Evan Sadler, George J. Broze, Joseph P. Miletich

Research output: Contribution to journalArticle

148 Scopus citations

Abstract

A human placenta cDNA library in λgtl 1 was screened for the expression of tissue factor antigens with rabbit polyclonal anti-human tissue factor immunoglobulin G. Among 4 million recombinant clones screened, one positive, XHTF8, expressed a protein that shared epitopes with authentic human brain tissue factor. The 1.1-kilobase cDNA insert of XHTF8 encoded a peptide that contained the amino-terminal protein sequence of human brain tissue factor. Northern blotting identified a major mRNA species of 2.2 kilobases and a minor species of ~3.2 kilobases in poly(A)+ RNA of placenta. Only 2.2-kilobase mRNA was detected in human brain and in the human monocytic U937 cell line. In U937 cells, the quantity of tissue factor mRNA was increased severalfold by exposure of the cells to phorbol 12-myristate 13-acetate. Additional cDNA clones were selected by hybridization with the cDNA insert of XHTF8. These overlapping isolates span 2177 base pairs of the tissue factor cDNA sequence that includes a 5’-noncoding region of 75 base pairs, an open reading frame of 885 base pairs, a stop codon, a 3’-noncoding region of 1141 base pairs, and a poly(A) tail. The open reading frame encodes a 33-kilodalton protein of 295 amino acids. The predicted sequence includes a signal peptide of 32 or 34 amino acids, a probable extracellular factor VII binding domain of 217 or 219 amino acids, a transmembrane segment of 23 amino acids, and a cytoplasmic tail of 21 amino acids. There are three potential glycosylation sites with the sequence Asn-X-Thr/Ser. The 3’-noncoding region contains an inverted Alu family repetitive sequence. The tissue factor gene was localized to chromosome 1 by hybridization of the cDNA insert of XHTF8 to flow-sorted human chromosomes.

Original languageEnglish
Pages (from-to)5234-5238
Number of pages5
JournalBiochemistry
Volume26
Issue number17
DOIs
StatePublished - Jan 1 1987

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    Scarpati, E. M., Flandermeyer, R. R., Siegel, N. R., Wen, D., Sadler, J. E., Broze, G. J., & Miletich, J. P. (1987). Human Tissue Factor: DNA Sequence and Chromosome Localization of the Gene. Biochemistry, 26(17), 5234-5238. https://doi.org/10.1021/bi00391a004