The murine platelet and plasma factor V pools are biosynthetically distinct and sufficient for minimal hemostasis

Blood ◽  
2003 ◽  
Vol 102 (8) ◽  
pp. 2856-2861 ◽  
Author(s):  
Hongmin Sun ◽  
Tony L. Yang ◽  
Angela Yang ◽  
Xixi Wang ◽  
David Ginsburg
Keyword(s):  
Gene Expression ◽  
Bacterial Artificial Chromosome ◽  
Transgenic Mice ◽  
Coagulation Factor ◽  
Artificial Chromosome ◽  
Coagulation Cascade ◽  
Factor V ◽  
Wild Type ◽  
Plasma Factor ◽  
Coagulation Factor V

Abstract Coagulation factor V (FV) is a central regulator of the coagulation cascade. Circulating FV is found in plasma and within platelet α granules. The specific functions of these distinct FV pools are uncertain. We now report the generation of transgenic mice with FV gene expression restricted to either the liver or megakaryocyte/platelet lineage using bacterial artificial chromosome (BAC) constructs. Six of 6 independent albumin BAC transgenes rescue the neonatal lethal hemorrhage of FV deficiency. Rescued mice all exhibit liver-specific Fv expression at levels ranging from 6% to 46% of the endogenous Fv gene, with no detectable FV activity within the platelet pool. One of the 3 Pf4 BAC transgenes available for analysis also rescues the lethal FV null phenotype, with FV activity restricted to only the platelet pool (approximately 3% of the wild-type FV level). FV-null mice rescued by either the albumin or Pf4 BAC exhibit nearly normal tail bleeding times. These results demonstrate that Fv expression in either the platelet or plasma FV pool is sufficient for basal hemostasis. In addition, these findings indicate that the murine platelet and plasma FV pools are biosynthetically distinct, in contrast to a previous report demonstrating a plasma origin for platelet FV in humans.

scholarly journals Activation of p90 Ribosomal S6 Kinases by ORF45 of Kaposi's Sarcoma-Associated Herpesvirus Is Critical for Optimal Production of Infectious Viruses

2014 ◽  
Vol 89 (1) ◽  
pp. 195-207 ◽  
Author(s):  
Bishi Fu ◽  
Ersheng Kuang ◽  
Wenwei Li ◽  
Denis Avey ◽  
Xiaojuan Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacterial Artificial Chromosome ◽  
Critical Role ◽  
Viral Gene Expression ◽  
Artificial Chromosome ◽  
Lytic Replication ◽  
Viral Gene ◽  
Wild Type ◽  
Lytic Reactivation ◽  
Infected Cells

ABSTRACTWe have previously shown that ORF45, an immediate-early and tegument protein of Kaposi's sarcoma-associated herpesvirus (KSHV), causes sustained activation of p90 ribosomal S6 kinases (RSKs) and extracellular regulated kinase (ERK) (E. Kuang, Q. Tang, G. G. Maul, and F. Zhu, J Virol 82:1838–1850, 2008,http://dx.doi.org/10.1128/JVI.02119-07). We now have identified the critical region of ORF45 that is involved in RSK interaction and activation. Alanine scanning mutagenesis of this region revealed that a single F66A point mutation abolished binding of ORF45 to RSK or ERK and, consequently, its ability to activate the kinases. We introduced the F66A mutation into BAC16 (a bacterial artificial chromosome clone containing the entire infectious KSHV genome), producing BAC16-45F66A. In parallel, we also repaired the mutation and obtained a revertant, BAC16-45A66F. The reconstitution of these mutants in iSLK cells demonstrated that the ORF45-F66A mutant failed to cause sustained ERK and RSK activation during lytic reactivation, resulting in dramatic differences in the phosphoproteomic profile between the wild-type virus-infected cells and the mutant virus-infected cells. ORF45 mutation or deletion also was accompanied by a noticeable decreased in viral gene expression during lytic reactivation. Consequently, the ORF45-F66A mutant produced significantly fewer infectious progeny virions than the wild type or the revertant. These results suggest a critical role for ORF45-mediated RSK activation in KSHV lytic replication.IMPORTANCEKSHV is the causative agent of three human malignancies. KSHV pathogenesis is intimately linked to its ability to modulate the host cell microenvironment and to facilitate efficient production of progeny viral particles. We previously described the mechanism by which the KSHV lytic protein ORF45 activates the cellular kinases ERK and RSK. We now have mapped the critical region of ORF45 responsible for binding and activation of ERK/RSK to a single residue, F66. We mutated this amino acid of ORF45 (F66A) and introduced the mutation into a newly developed bacterial artificial chromosome containing the KSHV genome (BAC16). This system has provided us with a useful tool to characterize the functions of ORF45-activated RSK upon KSHV lytic reactivation. We show that viral gene expression and virion production are significantly reduced by F66A mutation, indicating a critical role for ORF45-activated RSK during KSHV lytic replication.

Coagulation Factor V Leiden Mutation Was Detected in the Patients with Activated Protein C Resistance in Thailand

1998 ◽  
Vol 80 (08) ◽  
pp. 344-345 ◽  
Author(s):  
Pasra Arnutti ◽  
Motofumi Hiyoshi ◽  
Wichai Prayoonwiwat ◽  
Oytip Nathalang ◽  
Chamaiporn Suwanasophon ◽  
...  
Keyword(s):  
Protein C ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Coagulation Factor ◽  
Factor V ◽  
Activated Protein C Resistance ◽  
Factor V Leiden Mutation ◽  
Coagulation Factor V

Phenotyping and Genotyping of Coagulation Factor V Leiden

1996 ◽  
Vol 75 (02) ◽  
pp. 267-269 ◽  
Author(s):  
H Engel ◽  
L Zwang ◽  
H H D M van Vliet ◽  
J J Michiles ◽  
J Stibbe ◽  
...  
Keyword(s):  
Factor V Leiden ◽  
Coagulation Factor ◽  
Diagnostic Value ◽  
Resistance Test ◽  
Amplification Refractory Mutation System ◽  
Factor V ◽  
Chain Reactions ◽  
Apc Resistance ◽  
Specificity And Sensitivity ◽  
Coagulation Factor V

SummaryThe currently used activated Protein C resistance test demonstrated to be of limited diagnostic value for the detection of the mutant Factor V Leiden. Moreover, this assay is not useful for patients under anticoagulant therapy. A modification of the APC resistance test, applying Factor V deficient plasma is described which demonstrates a specificity and sensitivity of 1.0. The superiority of the modified APC resistance test over the existing APC resistance test was verified by genotyping.For that purpose, the Amplification Refractory Mutation System (ARMS) was applied to the detection of the G to A mutation at position 1691 in the gene encoding coagulation Factor V. The mutation at that position could be easily detected by using each of two allele-specific oligonucleotide primers concomitantly with one common primer in two separate polymerase chain reactions, thereby amplifying a fragment of 186 base-pairs of the Factor V gene.

Coagulation Factor V: An Old Star Shines Again

1997 ◽  
Vol 78 (01) ◽  
pp. 427-433 ◽  
Author(s):  
Jan Rosing ◽  
Guido Tans
Keyword(s):  
Coagulation Factor ◽  
Factor V ◽  
Coagulation Factor V

scholarly journals Association of coagulation factor V with the platelet cytoskeleton.

1982 ◽  
Vol 257 (8) ◽  
pp. 4557-4563
Author(s):  
G P Tuszynski ◽  
P N Walsh ◽  
J R Piperno ◽  
A Koshy
Keyword(s):  
Coagulation Factor ◽  
Factor V ◽  
Coagulation Factor V

scholarly journals Thrombin-catalyzed activation of human coagulation factor V.

1982 ◽  
Vol 257 (11) ◽  
pp. 6556-6564 ◽  
Author(s):  
K Suzuki ◽  
B Dahlbäck ◽  
J Stenflo
Keyword(s):  
Coagulation Factor ◽  
Factor V ◽  
Coagulation Factor V

scholarly journals Factor V Kuwait: A Novel Mutation in the Coagulation Factor V Gene Discovered in Kuwait

2006 ◽  
Vol 15 (2) ◽  
pp. 102-105
Author(s):  
Mehrez M. Jadaon ◽  
Ali A. Dashti ◽  
Hend L. Lewis
Keyword(s):  
Novel Mutation ◽  
Coagulation Factor ◽  
Factor V ◽  
Coagulation Factor V ◽  
Factor V Gene ◽  
V Gene
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