scholarly journals Correction of a dominant-negative von Willebrand factor multimerization defect by small interfering RNA-mediated allele-specific inhibition of mutant von Willebrand factor

2018 ◽  
Vol 16 (7) ◽  
pp. 1357-1368 ◽  
Author(s):  
A. de Jong ◽  
R. J. Dirven ◽  
J. A. Oud ◽  
D. Tio ◽  
B. J. M van Vlijmen ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Small Interfering Rna ◽  
Dominant Negative ◽  
Specific Inhibition ◽  
Allele Specific ◽  
Von Willebrand ◽  
Interfering Rna ◽  
Willebrand Factor

scholarly journals GH1 Splicing Is Regulated by Multiple Enhancers Whose Mutation Produces a Dominant-Negative GH Isoform That Can Be Degraded by Allele-Specific Small Interfering RNA (siRNA)

Endocrinology ◽  
2004 ◽  
Vol 145 (6) ◽  
pp. 2988-2996 ◽  
Author(s):  
Robin C. C. Ryther ◽  
Alex S. Flynt ◽  
Bryan D. Harris ◽  
John A. Phillips ◽  
James G. Patton
Keyword(s):  
Small Interfering Rna ◽  
Dominant Negative ◽  
Allele Specific ◽  
Interfering Rna

Three Distinct Candidate Point Mutations of the von Willebrand Factor Gene in Four Patients with Type IIA von Willebrand Disease

1992 ◽  
Vol 67 (06) ◽  
pp. 612-617 ◽  
Author(s):  
Isamu Sugiura ◽  
Tadashi Matsushita ◽  
Mitsune Tanimoto ◽  
Isao Takahashi ◽  
Tomio Yamazaki ◽  
...  
Keyword(s):  
Amino Acid ◽  
Von Willebrand Factor ◽  
Cleavage Site ◽  
Point Mutations ◽  
Pcr Amplification ◽  
Von Willebrand Disease ◽  
Missense Mutations ◽  
Allele Specific ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryType IIA von Willebrand disease (vWD) is the most common type II vWD and is characterized by the selective loss of large and intermediate sized multimers. One explanation for this disorder has been postulated to be a qualitative defect in von Willebrand factor (vWF) which results in increased susceptibility to proteolysis at the bond between residues Tyr842 and Met843. Four missense mutations that may cause type IIA vWD have recently been identified near the cleavage site. We analyzed the molecular basis for type IIA vWD in six patients. A 512 bp DNA sequence spanning the proteolytic cleavage site was targeted for PCR amplification and sequencing. We exploited a difference in restriction sites between the vWF gene and the pseudogene and have designed allele-specific oligomer used with PCR to distinguish these two genes. Three candidate missense mutations; Ser743 (TCG) → Leu (TTG), Leu799 (CTG) → Pro (CCG), and Arg834 (CGG) → Trp (TGG) were identified in 4 out of 6 patients. The amino acid substitution at Arg834 has been reported previously, but the other substitutions at Ser743 and Leu799 are novel candidate mutations locating 99 and 43 amino acids to the N-terminal side of the cleavage site, respectively. Our results indicate that amino acid substitutions located relatively distant from the cleavage site may also be involved in type IIA vWD.

ABO blood group genotypes, plasma von Willebrand factor levels and loading of von Willebrand factor with A and B antigens

2007 ◽  
Vol 97 (04) ◽  
pp. 534-541 ◽  
Author(s):  
Vania Morelli ◽  
Marieke de Visser ◽  
Nico van Tilburg ◽  
Hans Vos ◽  
Jeroen Eikenboom ◽  
...  
Keyword(s):  
Blood Group ◽  
Von Willebrand Factor ◽  
Case Control Study ◽  
Abo Blood Group ◽  
Allele Specific ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Control Study ◽  
Abo Genotypes ◽  
Large Case

SummaryABO blood group is a genetic determinant of von Willebrand factor (VWF) levels. We investigated the effect of ABO genotypes on VWF and factor VIII (FVIII) levels and on the degree to which VWF is loaded with A- and B-antigens, expressed as normalized ratios, nA-ratio and nB-ratio, respectively, in the Leiden Thrombophilia Study, a large case-control study on venous thrombosis. We found that the ABO locus had an allele-specific, dosage dependent effect on VWF and FVIII levels and on the loading of VWF with A-antigen and B-antigen. The highest mean nA- and nB-ratios were found in A 1 A 1 and BB genotypes, respectively. Four A1 O carriers had four 43-bp repeats in the minisatellite region of theABO gene in stead of the expected one repeat. All had a reduced nA-ratio compared to A 1 O carriers with one repeat in their A1 allele. The amount of A – and B- antigens expressed onVWF (nA-ratio and nB-ratio) explained about 18% (R2) of the variation in VWF levels.

scholarly journals Fluvastatin inhibits regulated secretion of endothelial cell von Willebrand factor in response to diverse secretagogues

2007 ◽  
Vol 405 (3) ◽  
pp. 597-604 ◽  
Author(s):  
Richard J. Fish ◽  
Hong Yang ◽  
Christelle Viglino ◽  
Raoul Schorer ◽  
Sylvie Dunoyer-Geindre ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Intracellular Calcium ◽  
Von Willebrand Factor ◽  
Small Gtpases ◽  
Dominant Negative ◽  
Calcium Handling ◽  
Regulated Secretion ◽  
Nos Inhibitors ◽  
Von Willebrand ◽  
Willebrand Factor

Regulated secretion of EC (endothelial cell) vWF (von Willebrand factor) is part of the haemostatic response. It occurs in response to secretagogues that raise intracellular calcium or cAMP. Statins are cholesterol-lowering drugs used for the treatment of cardiovascular disease. We studied the effect of fluvastatin on regulated secretion of vWF from HUVEC (human umbilical-vein ECs). Secretion in response to thrombin, a protease-activated receptor-1 agonist peptide, histamine, forskolin and adrenaline (epinephrine) was inhibited. This inhibition was reversed by mevalonate or geranylgeranyl pyrophosphate, and mimicked by a geranylgeranyl transferase inhibitor, demonstrating that the inhibitory mechanism includes inhibition of protein geranylgeranylation. To investigate this mechanism further, calcium handling and NO (nitric oxide) regulation were studied in fluvastatin-treated HUVEC. Intracellular calcium mobilization did not correlate with vWF secretion. Fluvastatin increased eNOS [endothelial NOS (NO synthase)] expression, but NOS inhibitors failed to reverse the effect of fluvastatin on vWF secretion. Exogenous NO did not inhibit thrombin-induced vWF secretion. Many small GTPases are geranylgeranylated and some are activated by secretagogues. We overexpressed DN (dominant negative) Rho GTPases, RhoA, Rac1 and Cdc42 (cell division cycle 42), in HUVEC. DNCdc42 conferred inhibition of thrombin- and forskolin-induced vWF secretion. We conclude that, via inhibition of protein geranylgeranylation, fluvastatin is a broadspectrum inhibitor of regulated vWF secretion. Geranylgeranylated small GTPases with functional roles in regulated secretion, such as Cdc42, are potential targets for the inhibitory activity of fluvastatin.

scholarly journals Plasma membrane phosphatidylinositol (4,5)-bisphosphate promotes Weibel–Palade body exocytosis

2020 ◽  
Vol 3 (11) ◽  
pp. e202000788
Author(s):  
Tu Thi Ngoc Nguyen ◽  
Sophia N Koerdt ◽  
Volker Gerke
Keyword(s):  
Plasma Membrane ◽  
Von Willebrand Factor ◽  
Membrane Lipids ◽  
Phospholipid Composition ◽  
Dominant Negative ◽  
Interaction Sites ◽  
Membrane Contact Sites ◽  
Phospholipase D1 ◽  
Von Willebrand ◽  
Willebrand Factor

Weibel–Palade bodies (WPB) are specialized secretory organelles of endothelial cells that control vascular hemostasis by regulated, Ca2+-dependent exocytosis of the coagulation-promoting von-Willebrand factor. Some proteins of the WPB docking and fusion machinery have been identified but a role of membrane lipids in regulated WPB exocytosis has so far remained elusive. We show here that the plasma membrane phospholipid composition affects Ca2+-dependent WPB exocytosis and von-Willebrand factor release. Phosphatidylinositol (4,5)-bisphosphate [PI(4,5)P2] becomes enriched at WPB–plasma membrane contact sites at the time of fusion, most likely downstream of phospholipase D1-mediated production of phosphatidic acid (PA) that activates phosphatidylinositol 4-phosphate (PI4P) 5-kinase γ. Depletion of plasma membrane PI(4,5)P2 or down-regulation of PI4P 5-kinase γ interferes with histamine-evoked and Ca2+-dependent WPB exocytosis and a mutant PI4P 5-kinase γ incapable of binding PA affects WPB exocytosis in a dominant-negative manner. This indicates that a unique PI(4,5)P2-rich environment in the plasma membrane governs WPB fusion possibly by providing interaction sites for WPB-associated docking factors.

The dominant-negative von Willebrand factor gene deletion p.P1127_C1948delinsR: molecular mechanism and modulation

Blood ◽  
2010 ◽  
Vol 116 (24) ◽  
pp. 5371-5376 ◽  
Author(s):  
Caterina Casari ◽  
Mirko Pinotti ◽  
Stefano Lancellotti ◽  
Elena Adinolfi ◽  
Alessandra Casonato ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Molecular Mechanisms ◽  
Von Willebrand Disease ◽  
Dominant Negative ◽  
Dominant Negative Effect ◽  
Wild Type ◽  
Collagen Binding ◽  
Cellular Models ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractUnderstanding molecular mechanisms in the dominant inheritance of von Willebrand disease would improve our knowledge of pathophysiologic processes underlying its prevalence. Cellular models of severe type 2 von Willebrand disease, caused by a heterozygous deletion in the von Willebrand factor (VWF) gene, were produced to investigate the altered biosynthesis. Coexpression of the wild-type and in-frame deleted (p.P1127_C1948delinsR) VWF forms impaired protein secretion, high molecular weight multimer formation and function (VWF collagen-binding 1.9% ± 0.5% of wild-type), which mimicked the patient's phenotype. mRNA, protein, and cellular studies delineated the highly efficient dominant-negative mechanism, based on the key role of heterodimers as multimer terminators. The altered VWF, synthesized in large amounts with the correctly encoded “cysteine knot” domain, formed heterodimers and heterotetramers with wild-type VWF, in addition to deleted homodimers. Impaired multimerization was associated with reduced amounts of VWF in late endosomes. Correction of the dominant-negative effect was explored by siRNAs targeting the mRNA breakpoint, which selectively inhibited the in-frame deleted VWF expression. Although the small amount of the deleted protein synthesized after inhibition still exerted dominant, even though weakened, negative effects, the siRNA treatment restored secretion of large multimers with improved function (VWF collagen-binding 28.0% ± 3.3% of wild-type).

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