Five novel mutations in the gene for human blood coagulation factor V associated with type I factor V deficiency

Blood ◽  
2001 ◽  
Vol 98 (2) ◽  
pp. 358-367 ◽  
Author(s):  
Richard van Wijk ◽  
Karel Nieuwenhuis ◽  
Marijke van den Berg ◽  
Eric G. Huizinga ◽  
Brenda B. van der Meijden ◽  
...  

Coagulation factor V (FV) plays an important role in maintaining the hemostatic balance in both the formation of thrombin in the procoagulant pathway as well as in the protein C anticoagulant pathway. FV deficiency is a rare bleeding disorder with variable phenotypic expression. Little is known about the molecular basis underlying this disease. This study identified 5 novel mutations associated with FV deficiency in 3 patients with severe FV deficiency but different clinical expression and 2 unaffected carriers. Four mutations led to a premature termination codon either by a nonsense mutation (single-letter amino acid codes): A1102T, K310Term. (FV Amersfoort) and C2491T, Q773Term. (FV Casablanca) or a frameshift: an 8–base pair deletion between nucleotides 1130 and 1139 (FV Seoul1) and a 1–base pair deletion between nucleotides 4291 and 4294 (FV Utrecht). One mutation was a novel missense mutation: T1927C, C585R (FV Nijkerk), resulting in the absence of mutant protein despite normal transcription to RNA. Most likely, an arginine at this position disrupts the hydrophobic interior of the FV A2 domain. The sixth detected mutation was a previously reported missense mutation: A5279G, Y1702C (FV Seoul2). In all cases, the presence of the mutation was associated with type I FV deficiency. Identifying the molecular basis of mutations underlying this rare coagulation disorder will help to obtain more insight into the mechanisms involved in the variable clinical phenotype of patients with FV deficiency.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1030-1030
Author(s):  
Rinku Patel ◽  
Jacky Cutler ◽  
Campbell Tait ◽  
Geoffrey Savidge

Abstract Factor V is a rare bleeding disorder characterized by low levels of plasma and/or platelet factor V, with an estimated prevalence of 1/1,000,000 and results from defects in the factor V gene. Human coagulation factor V (FV), a single chain glycoprotein, is synthesized in hepatocytes and megakaryocytes with 80% circulating free in plasma and the remainder being released upon platelet activation. The factor V gene comprises 25 exons ranging in size from 72bp to 2820bp coding for a protein which is oriented as A1-A2-B-A3-C1-C2 domains. FV is activated to its active form (FVa) by thrombin or activated factor X (FXa) which removes the B domain, generating a heavy chain and a light chain that are linked together in the presence of calcium ions. FVa binds to FXa and serves as its cofactor in the prothrombinase complex to convert prothrombin to thrombin. There is a high degree of homology between the A and C domains of FV and FVIII. We have investigated 8 unrelated patients from two centres with phenotypic and clinical charisteristics of FV deficiency. Mutation screening was carried out in these patients using Denaturing high performance liquid chromatography (dHPLC) and sequencing. Probable causative mutations were identified in all patients. A total of 10 novel mutations were identified in 8 patients and were located in the A1, A2, A3 and B domains. No mutations were identified in the C domain, and entries on the FV mutation database support our findings that mutations in this domain are less common than elsewhere in this gene. 5/8 patients were diagnosed with mild-moderate FV deficiency, and single heterozygous mutations were identified in each of these patients. 3 missense , 1 donor splicesite and 1 nonsense mutation were identified in the A1, A3 and B domains. The remaining 3/8 patients had severe FV deficiency (FV levels <2u/dl). One was compound heterozygous for 2 missense mutations in the A3 domain; one had a missense mutation in the A2 domain and a frameshift mutation (insertion of a single base pair) in the A3 domain. We have as yet identified only a heterozygous missense mutation in the third patient with severe FV deficiency. Phenotypic data and family history are strongly suggestive of the presence of a second mutation. Quantitative DNA analysis has confirmed the presence of 2 FV alleles, and RNA analysis is in progress to identify the second mutation. 100 normal alleles were analysed by dHPLC analysis or allele specific amplification to exclude these changes from being polymorphisms. We also have examined the homology between factor V and factor VIII, and the degree of similarity, between native and mutant amino acids to support these mutations as being causative of FV deficiency.


1999 ◽  
Vol 14 (4) ◽  
pp. 357-357 ◽  
Author(s):  
H. Kostka ◽  
G. Siegert ◽  
S. Gehrisch ◽  
E. Kuhlisch ◽  
E. Runge ◽  
...  

1996 ◽  
Vol 75 (03) ◽  
pp. 437-444 ◽  
Author(s):  
Jérôme Duchemin ◽  
Jeanne-Yvonne Borg ◽  
Delphine Borgel ◽  
Marc Vasse ◽  
Hervé Lévèque ◽  
...  

SummaryTo further elucidate the molecular basis for hereditary thrombophilia, we screened the protein S active gene in 11 families with type I deficiency, using a strategy based on denaturating gradient gel electrophoresis (DGGE) of all the coding sequences. Fragments with an abnormal DGGE pattern were sequenced, and 5 novel mutations were identified in 8 families. The mutations were a 7-nucleotide deletion in exon II, a 4-nucleotide deletion in exon III, a T insertion in exon VII, a C to T transition transforming Leu 259 into Pro and a T to C transition transforming Cys 625 into Arg in 4 families. These mutations were the only sequence variations found in the propositus’ gene exons and co-segregated with the plasma phenotype. A total of 28 members of these 8 families were heterozygous for one of the 5 mutations. Twenty-four (58,5%) of the 41 deficient subjects over 18 years of age had clinical thrombophilia, whereas the 13 subjects under 18 were asymptomatic. Of the 28 subjects, 6 (21,5%) were also found to bear the factor V Arg 506 Gin mutation.


1998 ◽  
Vol 80 (08) ◽  
pp. 344-345 ◽  
Author(s):  
Pasra Arnutti ◽  
Motofumi Hiyoshi ◽  
Wichai Prayoonwiwat ◽  
Oytip Nathalang ◽  
Chamaiporn Suwanasophon ◽  
...  

1996 ◽  
Vol 75 (02) ◽  
pp. 267-269 ◽  
Author(s):  
H Engel ◽  
L Zwang ◽  
H H D M van Vliet ◽  
J J Michiles ◽  
J Stibbe ◽  
...  

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.


Sign in / Sign up

Export Citation Format

Share Document