Mitogen-activated protein kinase regulates FSH-induced expression of tissue-type plasminogen activator through an activator protein 1 response element

This study was undertaken to investigate, in freshly isolated rat Sertoli cells, the physiological function of the type I and type II cyclic adenosine monophosphate (cAMP)-dependent protein kinase isozymes in tissue-type plasminogen activator secretion and the regulation of this cAMP process by follicle-stimulating hormone (FSH). Follicle-stimulating hormone-induced tissue-type plasminogen activator secretion depends upon intracellular cAMP levels. The changes in cAMP amounts required to activate maximally the tissue-type plasminogen activator secretion are extremely small, a cAMP threshold having to be reached for triggering the tissue-type plasminogen activator output. Intact Sertoli cells were incubated with combinations of cAMP analogs specific for each cAMP-dependent protein kinase type and complementary in their cAMP binding site on the cAMP-dependent protein kinase regulatory subunits: 8-aminohexylamino-cAMP = type 1, site 1; 8-thiomethyl-cAMP = type II, site 1 and N6-benzoyl-cAMP = types I/II, site 2. This allowed us to activate selectively each cAMP-dependent protein kinase type in a synergistic manner and then to evaluate their respective influence in the specific tissue-type plasminogen activator response. We establish that both of the cAMP-dependent protein kinase types are present and functional; the activity of the type I isozyme is preponderant (60%) in the cAMP-dependent tissue-type plasminogen activator secretion. Likewise, when these cAMP analogs were coupled with endogenously generated cAMP by FSH or forskolin, both of the cAMP-dependent protein kinase types were involved in the tissue-type plasminogen activator production. However, only tissue-type plasminogen activator secretion induced by FSH is mediated predominantly by the type I cAMP-dependent protein kinase, although the type II isozyme sustains an appreciable physiological role in the transmission pathway. We suggest some differences in the pattern of action between FSH and forskolin in Sertoli cells.

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Role of Distinct Mitogen-Activated Protein Kinase Pathways and Cooperation between Ets-2, ATF-2, and Jun Family Members in Human Urokinase-Type Plasminogen Activator Gene Induction by Interleukin-1 and Tetradecanoyl Phorbol Acetate

Molecular and Cellular Biology ◽

10.1128/mcb.19.9.6240 ◽

1999 ◽

Vol 19 (9) ◽

pp. 6240-6252 ◽

Cited By ~ 34

Author(s):

Grazia Cirillo ◽

Laura Casalino ◽

Daniela Vallone ◽

Anna Caracciolo ◽

Dario De Cesare ◽

...

Keyword(s):

Protein Kinase ◽

Plasminogen Activator ◽

Mutational Analysis ◽

Interleukin 1 ◽

Mitogen Activated Protein Kinase ◽

Transactivation Domain ◽

Tetradecanoyl Phorbol Acetate ◽

Type Plasminogen Activator ◽

Urokinase Type Plasminogen Activator ◽

Mitogen Activated Protein

ABSTRACT We have investigated the in vivo and in vitro regulation of the human urokinase-type plasminogen activator (uPA) gene by interleukin-1 (IL-1) and analyzed the transcription factors and signalling pathways involved in the response of the −2.0-kb uPA enhancer to IL-1 induction and to tetradecanoyl phorbol acetate (TPA) induction. Mutational analysis showed the cooperative activity of the Ets-binding site (EBS) and the two AP-1 elements of the enhancer. The results reveal that the EBS is required for the response to both inducers mediated by Ets-2, which is regulated at a level subsequent to DNA binding, by an IL-1- and phorbol ester-inducible transactivation domain. Both the IL-1 and the TPA-mediated induction result in a drastic increase of AP-1 binding to the downstream site of the enhancer (uPA 3′ TPA-responsive element), while a mostly qualitative change, resulting from the interplay between ATF-2 homodimers and c-Jun–ATF-2 heterodimers, takes place at the upstream AP-1 element. The analysis of two distinct mitogen-activated protein kinase pathways shows that stress-activated protein kinase–Jun N-terminal kinase activation, resulting in the phosphorylation of ATF-2, c-Jun, and JunD, is required not only for the IL-1- but also for the TPA-dependent induction, while the extracellular signal-related kinase 1 (ERK-1) and ERK-2 activation is involved in the TPA- but not in the IL-1-dependent stimulation of the uPA enhancer.

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Inhibition of coagulation, fibrinolysis, and endothelial cell activation by a p38 mitogen-activated protein kinase inhibitor during human endotoxemia

Blood ◽

10.1182/blood-2002-11-3338 ◽

2003 ◽

Vol 101 (11) ◽

pp. 4446-4448 ◽

Cited By ~ 51

Author(s):

Judith Branger ◽

Bernt van den Blink ◽

Sebastiaan Weijer ◽

Abhya Gupta ◽

Sander J.H. van Deventer ◽

...

Keyword(s):

Endothelial Cell ◽

Protein Kinase ◽

Plasminogen Activator ◽

P38 Mapk ◽

Cell Activation ◽

Plasma Concentrations ◽

Mitogen Activated Protein Kinase ◽

Tissue Type ◽

Endothelial Cell Activation ◽

Mitogen Activated Protein

Abstract P38 mitogen-activated protein kinase (MAPK) is an important component of intracellular signaling cascades that initiate various inflammatory cellular responses. To determine the role of p38 MAPK in the procoagulant response to lipopolysaccharide (LPS), 24 healthy subjects were exposed to an intravenous dose of LPS (4 ng/kg), preceded 3 hours earlier by orally administered 600 or 50 mg BIRB 796 BS (a specific p38 MAPK inhibitor), or placebo. The 600-mg dose of BIRB 796 BS strongly inhibited LPS-induced coagulation activation, as measured by plasma concentrations of the prothrombin fragment F1 + 2. BIRB 796 BS also dose dependently attenuated the activation and subsequent inhibition of the fibrinolytic system (plasma tissue-type plasminogen activator, plasmin-α2-antiplasmin complexes, and plasminogen activator inhibitor type 1) and endothelial cell activation (plasma soluble E-selectin and von Willebrand factor). Activation of p38 MAPK plays an important role in the procoagulant and endothelial cell response after in vivo exposure to LPS.

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P1705 Increase in venous tissue-type plasminogen activator (tPA) by exercise predicts local endothelial tPA release in vivo in healthy men

European Heart Journal ◽

10.1016/s0195-668x(03)94843-3 ◽

2003 ◽

Vol 24 (5) ◽

pp. 322

Author(s):

T GUDNASON

Keyword(s):

Plasminogen Activator ◽

Tissue Type ◽

Tissue Type Plasminogen Activator ◽

Healthy Men ◽

Type Plasminogen Activator ◽

Venous Tissue

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Binding of 131I-Labeled Tissue-Type Plasminogen Activator on De-Endothelialized Lesions in Rabbits

Nuklearmedizin ◽

10.1055/s-0038-1628893 ◽

1987 ◽

Vol 26 (05) ◽

pp. 224-228 ◽

Cited By ~ 2

Author(s):

Y. Isaka ◽

H. Etani ◽

K. Kimura ◽

S. Yoneda ◽

T. Kamada ◽

...

Keyword(s):

Plasminogen Activator ◽

Abdominal Aorta ◽

Half Life ◽

Balloon Catheter ◽

Biochemical Properties ◽

Tissue Type ◽

Fibrin Deposition ◽

Tissue Type Plasminogen Activator ◽

High Affinity ◽

Type Plasminogen Activator

Tissue-type plasminogen activator (t-PA) which has a high affinity for fibrin in the clot, was labeled with 131I by the iodogen method, and its binding to de-endothelialized lesions in the rabbit was measured to assess the detectability of thrombi. The de-endothelialized lesion was induced in the abdominal aorta with a Fogarty 4F balloon catheter. Two hours after the de-endothelialization, 131I-labeled t-PA (125 ± 46 μCi) was injected intravenously. The initial half-life of the agent in blood (n = 12) was 2.9 ± 0.4 min. The degree of binding of 131I-labeled t-PA to the de-endothelialized lesion was evaluated at 15 min (n = 6) or at 30 min (n = 6) after injection of the agent. In spite of the retention of the biochemical properties of 131I-labeled t-PA and the presence of fibrin deposition at the de-endothelialized lesion, the binding of t-PA to the lesion was not sufficiently strong. Lesion-to-control ratios (cpm/g/cpm/g) were 1.65 ± 0.40 (at 15 min) and 1.39 ± 1.31 (at 30 min), and lesion-to-blood ratios were 1.39 ± 0.32 (at 15 min) and 1.36 ± 0.23 (at 30 min). These results suggest that radiolabeled t-PA may be inappropriate as a radiopharmaceutical for the scintigraphic detection of a pre-existing thrombotic lesion.

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The Influence of Carbohydrate Structure on the Clearance of Recombinant Tissue-Type Plasminogen Activator

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647041 ◽

1988 ◽

Vol 60 (02) ◽

pp. 255-261 ◽

Cited By ~ 81

Author(s):

A Hotchkiss ◽

C J Refino ◽

C K Leonard ◽

J V O'Connor ◽

C Crowley ◽

...

Keyword(s):

Amino Acid ◽

Plasminogen Activator ◽

Sodium Periodate ◽

Glycosylation Site ◽

Tissue Type ◽

Carbohydrate Structure ◽

Tissue Type Plasminogen Activator ◽

Type Plasminogen Activator ◽

High Mannose ◽

Oligosaccharide Structures

SummaryModification of the carbohydrate structures of recombinant tissue-type plasminogen activator (rt-PA) can increase or decrease its rate of clearance in rabbits. When rt-PA was treated with sodium periodate to oxidize carbohydrate residues, the rate of clearance was decreased from 9.6 ± 1.9 ml min−1 kg−1 to 3.5 ± 0.6 ml min−1 kg−1 (mean ± SD, n = 5). A similar change in the clearance of rt-PA was introduced by the use of endo-β-N-acetyl- glucosaminidase H (Endo-H), which selectively removes high mannose asparagine-linked oligosaccharides; the clearance of Endo-H-treated rt-PA was 5.0 ± 0.5 ml min−1 kg−1. A mutant of rt-PA was produced with an amino acid substitution at position 117 (Asn replaced with Gin) to remove a potential glycosylation site that normally contains a high mannose structure. The clearance of this material was also decreased, similar to the periodate and Endo-H-treated rt-PA. Conversely, when rt-PA was produced in the CHO 15B cell line, which can produce only high mannose oligosaccharide structures on glycoproteins, the clearance was increased by a factor of 1.8. These results demonstrate that the removal of rt-PA from the blood depends significantly upon the nature of its oligosaccharide structures.

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