Functional characterization of a novel missense mutation identified in a Turkish patient affected by severe coagulation factor V deficiency

E. M. Paraboschi, S. M. Kayiran, N. Özbek, B. Gürakan, F. Peyvandi, I. Guella, S. Duga, R. Asselta

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Factor V (FV) deficiency is a rare coagulation disorder, characterized by a bleeding phenotype varying from mild to severe. To date, 115 mutations have been described along the gene encoding for FV (F5) but only few of them have been functionally characterized. Aim of this study was the identification and the molecular characterization of genetic defects underlying severe FV deficiency in a 7-month-old Turkish patient. Mutation detection was performed by sequencing the whole F5 coding region, exon-intron boundaries and about 300bp of the promoter region. Functional analysis of the identified missense mutation was conducted by transient expression of wild-type and mutant FV recombinant molecules in COS-1 cells. Two novel mutations: a missense (Pro132Arg) and a 1-bp deletion (Ile1890TyrfsX19) were identified in the F5 gene. While the frameshift mutation is responsible for the introduction of a premature stop codon, likely triggering F5 mRNA to nonsense-mediated mRNA degradation, the demonstration of the pathogenic role of the Pro132Arg mutation required an experimental validation. Expression experiments showed that the missense mutation causes a significant reduction in FV secretion and in the specific activity of the residual secreted molecule (77% and 78% decrease, respectively). This paper reports the identification of two novel mutations responsible for FV deficiency, thus widening the mutational spectrum of the F5 gene. The Pro132Arg mutation adds to the only other two functionally characterized missense defects in the FV A1 domain.

Original languageEnglish
Pages (from-to)205-210
Number of pages6
JournalHaemophilia
Volume18
Issue number2
DOIs
Publication statusPublished - Mar 2012

Fingerprint

Factor V Deficiency
Factor V
Missense Mutation
Mutation
Genes
Frameshift Mutation
Nonsense Codon
COS Cells
RNA Stability
Genetic Promoter Regions
Introns
Molecular Biology
Exons
Hemorrhage
Phenotype
Messenger RNA

Keywords

  • A1 domain
  • Coagulation factor V
  • Factor V deficiency
  • Frameshift mutation
  • In vitro expression
  • Missense mutation

ASJC Scopus subject areas

  • Hematology
  • Genetics(clinical)

Cite this

Functional characterization of a novel missense mutation identified in a Turkish patient affected by severe coagulation factor V deficiency. / Paraboschi, E. M.; Kayiran, S. M.; Özbek, N.; Gürakan, B.; Peyvandi, F.; Guella, I.; Duga, S.; Asselta, R.

In: Haemophilia, Vol. 18, No. 2, 03.2012, p. 205-210.

Research output: Contribution to journalArticle

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