scholarly journals Plasminogen activator inhibitor-1 promoter activity in adipocytes is not influenced by the 4 G/5 G promoter polymorphism and is regulated by a USF-1/2 binding site immediately preceding the polymorphic region

2004 ◽  
Vol 32 (1) ◽  
pp. 155-163 ◽  
Author(s):  
B Zietz ◽  
W Drobnik ◽  
H Herfarth ◽  
C Buechler ◽  
J Scholmerich ◽  
...  
Keyword(s):  
Binding Site ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Promoter Polymorphism ◽  
Luciferase Reporter ◽  
Plasminogen Activator Inhibitor 1 ◽  
Luciferase Reporter Gene ◽  
Polymorphic Region ◽  
Activator Inhibitor ◽  
Pai 1

Plasminogen activator inhibitor-1 (PAI-1) levels were found to be associated with obesity indicating that adipocytes influence PAI-1 plasma levels. In addition, the 4 G/5 G promoter polymorphism of the PAI-1 gene may modulate PAI-1 transcription. We investigated the transcriptional regulation of the human PAI-1 gene in adipocytes and analyzed the genetic contribution of the 4 G/5 G polymorphism. The PAI-1 promoter was analyzed using electrophoretic mobility shift assays (EMSAs) and luciferase reporter gene assays. A putative binding site for the upstream stimulatory factor-1/2 (USF-1/2) at the polymorphic region of the PAI-1 promoter was identified. The binding of USF-1/2 was studied using nuclear extracts prepared from adipocytes and was similar in all the promoter variants as analyzed by EMSA. A 257 bp PAI-1 promoter fragment including the 4 G/5 G site was transcriptionally active in adipocytes and was not influenced by the polymorphism. The present data indicate for the first time that USF-1/2 is transcriptionally active in differentiated adipocytes. However, USF-1/2 binding activity and PAI-1 transcription are not influenced by the 4 G/5 G-allele. These data possibly explain the observation that PAI-1 secretion from adipose tissue is not influenced by the PAI-1 promoter polymorphism.

Localization of a Vitronectin Binding Region of Plasminogen Activator Inhibitor-1

1995 ◽  
Vol 73 (05) ◽  
pp. 829-834 ◽  
Author(s):  
Jaya Padmanabhan ◽  
David C Sane
Keyword(s):  
Binding Site ◽  
Plasminogen Activator ◽  
Inhibitory Activity ◽  
Plasminogen Activator Inhibitor ◽  
Structural Homology ◽  
Binding Region ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor ◽  
Pai 1

SummaryThe PAI-1 binding site for VN was studied using two independent methods. PAI-1 was cleaved by Staph V8 protease, producing 8 fragments, only 2 of which bound to [125I]-VN. These fragments were predicted to overlap between residues 91-130. Since PAI-2 has structural homology to PAI-1, but does not bind to vitronectin, chimeras of PAI-1 and PAI-2 were constructed. Four chimeras, containing PAI-1 residues 1-70,1-105,1-114, and 1-167 were constructed and expressed in vitro. PAI-1, PAI-2, and all of the chimeras retained inhibitory activity for t-PA, but only the chimera containing PAI-1 residues 1-167 formed a complex with VN. Together, these results predict that the VN binding site of PAI-1 is between residues 115-130.

Plasminogen activator inhibitor-1 4G/5G promoter polymorphism and PAI-1 plasma levels in young patients with ischemic stroke

2011 ◽  
Vol 38 (8) ◽  
pp. 5355-5360 ◽  
Author(s):  
Adriano de Paula Sabino ◽  
Daniel Dias Ribeiro ◽  
Caroline Pereira Domingueti ◽  
Mariana Silva dos Santos ◽  
Telma Gadelha ◽  
...  
Keyword(s):  
Ischemic Stroke ◽  
Plasminogen Activator ◽  
Plasma Levels ◽  
Young Patients ◽  
Plasminogen Activator Inhibitor ◽  
Promoter Polymorphism ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor ◽  
Pai 1

scholarly journals Involvement of miR-30c and miR-301a in immediate induction of plasminogen activator inhibitor-1 by placental growth factor in human pulmonary endothelial cells

2011 ◽  
Vol 434 (3) ◽  
pp. 473-482 ◽  
Author(s):  
Nitin Patel ◽  
Stanley M. Tahara ◽  
Punam Malik ◽  
Vijay K. Kalra
Keyword(s):  
Endothelial Cells ◽  
Growth Factor ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Placental Growth Factor ◽  
Luciferase Reporter ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor ◽  
Pai 1

PAI-1 (plasminogen activator inhibitor-1) is a key physiological inhibitor of fibrinolysis. Previously, we have reported PlGF (placental growth factor)-mediated transcriptional up-regulation of PAI-1 (SERPINE1) mRNA expression via activation of HIF-1α (hypoxia-inducible factor-1α) and AP-1 (activator protein-1) in HPMVECs (human pulmonary microvascular endothelial cells), which resulted in elevated PAI-1 in humans with SCA (sickle cell anaemia). In the present study, we have identified the role of post-transcriptional mechanism(s) of PlGF-mediated accumulation of PAI-1 mRNA in HPMVECs by examining the role of microRNAs (miRNAs/miRs) in PlGF-induced PAI-1 mRNA stability. Our results show reduced expression of miR-30c and miR-301a, but not of miR-99a, in response to PlGF, which have evolutionarily conserved binding sites in the 3′-UTR (3′-untranslated region) of PAI-1 mRNA. Transfection of anti-miR-30c or anti-miR-301a oligonucleotides resulted in increased PAI-1 mRNA levels, which were increased further with PlGF stimulation. Conversely, overexpression of pre-miR-30c or pre-miR-301a resulted in an attenuation of PlGF-induced PAI-1 mRNA and protein levels. Luciferase reporter assays using wild-type and mutant 3′-UTR constructs confirmed that the PAI-1 3′-UTR is indeed a direct target of miR-30c and miR-301a. Finally, plasma levels of miR-30c and miR-301a were significantly down-regulated in patients with SCA compared with normal controls. These results provide a post-transcriptional regulatory mechanism of PlGF-induced PAI-1 elevation.

scholarly journals The role of plasminogen activator inhibitor-1 in gastric mucosal protection

2013 ◽  
Vol 304 (9) ◽  
pp. G814-G822 ◽  
Author(s):  
Susan Kenny ◽  
Islay Steele ◽  
Suzanne Lyons ◽  
Andrew R. Moore ◽  
Senthil V. Murugesan ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Luciferase Reporter ◽  
Gastric Epithelial Cells ◽  
Plasminogen Activator Inhibitor 1 ◽  
Luciferase Reporter Construct ◽  
Plasminogen Activator System ◽  
Activator Inhibitor ◽  
Pai 1

Gastric mucosal health is maintained in response to potentially damaging luminal factors. Aspirin and nonsteroidal anti-inflammatory drugs (NSAIDs) disrupt protective mechanisms leading to bleeding and ulceration. The plasminogen activator system has been implicated in fibrinolysis following gastric ulceration, and an inhibitor of this system, plasminogen activator inhibitor (PAI)-1, is expressed in gastric epithelial cells. In Helicobacter pylori-negative patients with normal gastric histology taking aspirin or NSAIDs, we found elevated gastric PAI-1 mRNA abundance compared with controls; the increase in patients on aspirin was independent of whether they were also taking proton pump inhibitors. In the same patients, aspirin tended to lower urokinase plasminogen activator mRNA. Immunohistochemistry indicated PAI-1 localization to epithelial cells. In a model system using MKN45 or AGS-GR cells transfected with a PAI-1 promoter-luciferase reporter construct, we found no evidence for upregulation of PAI-1 expression by indomethacin, and, in fact, cyclooxygenase products such as PGE2 and PGI2 weakly stimulated expression. Increased gastric PAI-1 mRNA was also found in mice following gavage with ethanol or indomethacin, but plasma PAI-1 was unaffected. In PAI-1−/− mice, gastric hemorrhagic lesions in response to ethanol or indomethacin were increased compared with C57BL/6 mice. In contrast, in PAI-1-H/Kβ mice in which PAI-1 is overexpressed in parietal cells, there were decreased lesions in response to ethanol and indomethacin. Thus, PAI-1 expression is increased in gastric epithelial cells in response to mucosal irritants such as aspirin and NSAIDs probably via an indirect mechanism, and PAI-1 acts as a local autoregulator to minimize mucosal damage.

Abstract 174: miR30c Regulates Plasminogen Activator Inhibitor-1 In Platelet

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Mao Luo ◽  
Qin Wan ◽  
Fei Liu ◽  
Rong Li ◽  
Jiyi Xia ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Protein Level ◽  
Target Gene ◽  
Plasminogen Activator Inhibitor ◽  
Human Platelets ◽  
Luciferase Reporter ◽  
Plasminogen Activator Inhibitor 1 ◽  
Gene Assay ◽  
Activator Inhibitor ◽  
Pai 1

Objective. PAI-1 mRNA and protein have been detected in human platelets. Recently some miRNAs have been found in human platelets, which are involved in the regulation of genes and the protein synthesis. However, little is known about the physiological roles of individual miRNAs in platelets. In this study, we investigated whether miR30c can regulate platelet-derived plasminogen activator inhibitor-1(PAI-1). Methods and Results. Expression of miR-30c, PAI-1, miR-21 and its targeted gene TIMP1 were found in healthy human leukocyte-depleted platelets (LDPs) by real time PCR. In luciferase reporter gene assay, miR-30c targets the 3’ untranslated region (3’ UTR) of PAI-1 mRNA through a miR-30c binding site. Transfection of miR-30c mimic into MEG-01, a megakaryoblastic cell line, significantly reduced PAI-1 protein level compared with negative control. Inhibition of miR-30c by transfecting miR-30c inhibitor significantly increased PAI-1 protein level. Furthermore, miR-21 expression was significantly down-regulated after transfecting with miR-30c mimic in PAI-1-/- mice LDPs, conversely, the expression of its target gene TIMP1 was significantly up-regulated after transfecting with miR-30c mimic in PAI-1-/- mice LDPs. Conclusion. These results provide a novel regulatory mechanism of miR30c- regulated PAI-1 protein through its influence on the downstream miR21 and its target gene TIMP1 expression in platelet, suggesting that miR-30c might be a potential new strategy for anti-thrombosis.

Role of 4G/5G promoter polymorphism of Plasminogen Activator Inhibitor-1 (PAI-1) gene in outcome of sepsis

2010 ◽  
Vol 125 (4) ◽  
pp. 367-369 ◽  
Author(s):  
Silvia Perés Wingeyer ◽  
Gabriela de Larrañaga ◽  
Eleonora Cunto ◽  
Laura Fontana ◽  
Cristina Nogueras ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Promoter Polymorphism ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor ◽  
Pai 1
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