Genomic structure and promoter activity of the human tissue factor pathway inhibitor-2 gene.
Human type-2 tissue factor pathway inhibitor (TFPI-2), also known as placental protein 5, is a 32 kDa serine proteinase inhibitor consisting of three tandemly arranged Kunitz-type inhibitor domains homologous to tissue factor pathway inhibitor. TFPI-2 strongly inhibits a wide variety of serine proteinases including trypsin, chymotrypsin, plasmin, kallikrein and blood coagulation factor XIa. In this study, we have isolated and characterized a genomic clone from an artificial chromosome genomic library that encodes the entire human TFPI-2 gene. The human TFPI-2 gene spans approximately 7 kb and consists of five exons and four introns. Each Kunitz-type domain is encoded by a single exon, similar to that observed for murine TFPI-2 and other Kunitz-type proteinase inhibitors. A total of 535 bp of the 3'-flanking region contain two probable polyadenylation sites (AATAAA) at +4297 and +4314. A single transcription initiation site was identified by oligo-capping and reverse transcription-PCR analysis. Transient transfection of reporter plasmids containing segments of the 5'-flanking region into human transformed bone marrow endothelial cells and glioblastoma cells identified an 85 bp region (-224 to -139) sufficient for transcription of the human TFPI-2 gene.
Kamei
S
S
Department of Pathology, University of New Mexico Health Science Center, Albuquerque 87131, USA.
Kazama
Y
Y
Kuijper
J L
JL
Foster
D C
DC
Kisiel
W
W
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005091:Exons; D015870:Gene Expression; D015894:Genome, Human; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D011257:Pregnancy Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D017384:Sequence Deletion; D015842:Serine Proteinase Inhibitors / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D006023:Glycoproteins; D011257:Pregnancy Proteins; D015842:Serine Proteinase Inhibitors; D014157:Transcription Factors; C021240:tissue-factor-pathway inhibitor 2