The human urokinase-plasminogen activator gene and its promoter

Abstract

The urokinase type of plasminogen activator (uPA) is subject to regulation by hormones, phorbol esters and oncogenic transformation. This enzyme has been suggested to play a key role in processes involving cell migration and tissue remodeling, and to be essential for tumor metastasis. In order to study these processes, we have isolated the human uPA gene, and have determined its entire nucleotide sequence. The gene is organized in 11 exons and is 6.4 kb long. The 5' end of uPA mRNA has been determined by both S1 mapping and primer extension experiments. A fragment of 800 bp containing the entire 5' flanking region shows promoter activity when introduced upstream of a bacterial chloramphenicol acetyltransferase gene and introduced into human cells. The hexanucleotide sequence GGCGGG, previously found at similar regions in several viral and eukaryotic promoters and shown to be essential for promoter activity (McKnight et al. (1984) Cell, 37, 253-262), is repeated three times between the CAAT and the TATA boxes.

Advanced glycation endproducts inhibit prostacyclin production and induce plasminogen activator inhibitor-1 in human microvascular endothelial cells

“…Oligonucleotide primers and probes for quantitative reverse transcription-polymerase chain reactions (RT-PCR) were chemically synthesized and purified as described previously [21]. Sequences of the upstream and downstream primers and the internal probe were 5¢-ATGGGATT-CAAGATTGATGA-3¢, 5¢-TCAGTATAGTTGAACTTG-TT-3¢ and 5¢-AGAGAGCCAGATTCATCATCAAT-3¢ (nucleotides (nt) 379±398, 811±830 and 584±606) for detecting PAI-1 mRNA (22), 5¢-ACCACCATCGAGAACCAGCC-3¢, 5¢-AATCAGCTTCACAACAGTCA-3¢ and 5¢-CTGGTTT-GCGGCCATCTACA-3¢ (nt 2494±2513, 4174±4193 and 2514± 2533) for u-PA mRNA [23], 5¢-CGAAGGATTTGCTGGG-AAGT-3¢, 5¢-TGCGGTTCTTCAGCACGTGG-3¢ and 5¢-TA-CGAGGACCAGGGCATCAG-3¢, (nt 245±264, 746±765 and 294±313) for tissue-plasminogen activator (t-PA) mRNA [24]. The primers and probe used for beta-actin mRNA detection were the same as described previously [21].…”

Osteopontin induces increased invasiveness and plasminogen activator expression of human mammary epithelial cells

“…Probes for human proteinase and uPAR genes included: MMP-9 (92 kDa Type IV collagenase) (1046 bp insert from plasmid p92MO1) (gift of Dr WG StetlerStevenson), MMP-2 (72 kDa Type IV collagenase) (1117 bp insert from plasmid p3Ha) (Reponen et al, 1992), cathepsin B (1.6 kb KpnI insert from plasmid pLC343) (gift of Dr B Sloane) (Cao et al, 1994), cathepsin D (20 kb insert from plasmid pM13mp10) (gift of Dr H Rochefort) (Augereau et al, 1988), cathepsin L (800 bp insert from plasmid pHCL800.1) (gift of Dr DT Denhardt) (Joseph et al, 1988), urokinase-type plasminogen activator (40mer antisense oligonucleotide derived from the translated sequences of exon 4) (Calbiochem/Cederlane Laboratories, Hornby, Ontario, Canada, Cat#ON333) (Riccio et al, 1985), urokinase-type plasminogen activator receptor (45mer antisense oligonucleotide probe to the ®rst 15 amino acids (not including the signal peptide) (Roldan et al, 1990). Even loading of lanes was con®rmed by probing blots with a human 18S rRNA probe (p100D9; a kind gift from Dr DT Denhardt).…”

In vitro inhibition of human malignant brain tumour cell line proliferation by anti-urokinase-type plasminogen activator monoclonal antibodies

“…Plasminogen actix ator is a serine protease existing in two forms known as tissue type (tPA) and urokinase tx-pe (uPA) (Sobel et al 1952: Giinzler et al 1982a.b: Nielsen et al 1982: Wun et al 1982: Salerno et al 1984: Riccio et al 1985. uPA converts plasminogen into plasmin and thus mediates pericellular proteolysis during cell migration and tissue remodelling, under phy siological and pathophy siological conditions (Gnrmann et al 1976: Gerdin and Saldeen.…”