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Human tissue factor: cDNA sequence and chromosome localization of the gene
E M Scarpati1, D Wen, G J Broze
1Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri 63110.
Biochemistry
|August 25, 1987
Summary
Researchers identified a human placenta cDNA clone expressing tissue factor, crucial for blood clotting. This discovery aids in understanding tissue factor gene expression and its role in various tissues.
Area of Science:
- Molecular Biology
- Immunology
- Biochemistry
Background:
- Tissue factor is a key initiator of the coagulation cascade.
- Understanding tissue factor gene expression is vital for hemostasis research.
Purpose of the Study:
- To isolate and characterize the cDNA encoding human tissue factor.
- To investigate tissue factor mRNA expression in different human tissues and cell lines.
Main Methods:
- Screening of a human placenta cDNA library using anti-human tissue factor antibodies.
- Northern blotting to analyze mRNA species.
- cDNA sequencing to determine the full gene sequence and protein structure.
Main Results:
- Isolation of a positive clone (lambda HTF8) expressing a protein with tissue factor epitopes.
- Identification of a 2.2 kb mRNA species in placenta, brain, and U937 cells, with a minor 3.2 kb species in placenta.
- Phorbol ester treatment significantly increased tissue factor mRNA in U937 cells.
- The complete cDNA sequence revealed an open reading frame encoding a 33 kDa protein with predicted signal peptide, extracellular domain, transmembrane segment, and cytoplasmic tail.
Conclusions:
- The isolated cDNA encodes human tissue factor, confirming its expression in placenta.
- Tissue factor mRNA is present in brain and monocytic cells, with differential expression regulation.
- The predicted protein structure provides insights into tissue factor's function and localization.
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