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Thromb Haemost 2008; 99(03): 523-530
DOI: 10.1160/TH07-12-0723
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Molecular characterization of two novel mutations causing factor XI deficiency: A splicing defect and a missense mutation responsible for a CRM+ defect

Ilaria Guella
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
,
Giulia Soldà
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
,
Silvia Spena
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
,
Rosanna Asselta
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
,
Rossella Ghiotto
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
,
Maria Luisa Tenchini
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
,
Giancarlo Castaman
2   Department of Hematology and Hemophilia and Thrombosis Center, San Bortolo Hospital, Vicenza, Italy
,
Stefano Duga
1   Department of Biology and Genetics for Medical Sciences, University of Milan, Milan, Italy
› Author Affiliations Financial support: S. Duga is a recipient of a Bayer Hemophilia Early Career Investigator Award 2006. This work was also supported by MIUR (Ministero dell’Istruzione, dell’Università e della Ricerca): FIRST 2006 (Fondo per gli Investimenti Ricerca Scientifica e Tecnologica).
Further Information

Publication History

Received: 06 December 2007

Accepted after major revision: 26 January 2008

Publication Date:
07 December 2017 (online)

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Summary

Severe factor XI (FXI) deficiency is a bleeding disorder generally inherited as an autosomal recessive trait and characterized by haemorrhagic symptoms mainly associated with injury or surgery. So far, more than 150 causative molecular defects have been identified throughout the F11 gene. In the present study, we investigated the molecular basis of FXI deficiency in two Italian patients. Mutational screening of the F|| gene disclosed a novel missense substitution (Arg184Gly) in exon 7 and two splicing mutations: a novel G>A transition affecting the last nucleotide of exon 4 (325G>A), and the already known IVS6+3A>G. RT-PCR assays were performed on total RNA extracted from platelets and lymphocytes of each patient. Se- quencing of RT-PCR products demonstrated that both 325G>A and IVS6+3A>G mutations abolish the corresponding donor splice site, causing the skipping of the affected exon; this in turn results in a frameshift introducing a premature termination codon. Expression of recombinant FXI-Arg184Gly revealed a 70% reduction in FXI activity, suggesting that the Arg184Gly mutation might cause a cross-reactive material positive (CRM+) deficiency. In conclusion, the functional consequences of two splicing mutations leading to FXI deficiency have been elucidated. Moreover, we report a novel missense mutation in the FIX-binding region of the FXI A3 domain leading to a CRM+ deficiency.

Keywords

FXI deficiency - mutational screening - expression experiments - splicing mutations - CRM+ defects
 

  • References

  • 1 Seligsohn U. Factor XI in haemostasis and thrombosis: past, present and future. Thromb Haemost 2007; 98: 84-89.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Gailani D, Broze GJ. Jr. Factor XI activation in a revised model of blood coagulation. Science 1991; 253: 909-912.
    CrossrefPubMedGoogle Scholar
  • 3 Bouma BN, Griffin JH. Human blood coagulation factor XI. Purification, properties, and mechanism of activation by activated factor XII. J Biol Chem 1977; 252: 6432-6437.
    PubMedGoogle Scholar
  • 4 Fujikawa K, Chung DW, Hendrickson LE. et al. Amino acid sequence of human factor XI, a blood coagulation factor with four tandem repeats that are highly homologous with plasma prekallikrein. Biochemistry 1986; 25: 2417-2424.
    CrossrefPubMedGoogle Scholar
  • 5 Baglia FA, Badellino KO, Ho DH. et al. A binding site for the kringle II domain of prothrombin in the apple 1 domain of factor XI. J Biol Chem 2000; 275: 31954-31962.
    CrossrefPubMedGoogle Scholar
  • 6 Baglia FA, Jameson BA, Walsh PN. Identification and chemical synthesis of a substrate-binding site for factor IX on coagulation factor XIa. J Biol Chem 1991; 266: 24190-24197.
    PubMedGoogle Scholar
  • 7 Baglia FA, Jameson BA, Walsh PN. Identification and characterization of a binding site for platelets in the Apple 3 domain of coagulation factor XI. J Biol Chem 1995; 270: 6734-6740.
    CrossrefPubMedGoogle Scholar
  • 8 Sun MF, Zhao M, Gailani D. Identification of amino acids in the factor XI apple 3 domain required for activation of factor IX. J Biol Chem 1999; 274: 36373-36378.
    CrossrefPubMedGoogle Scholar
  • 9 Baglia FA, Jameson BA, Walsh PN. Identification and characterization of a binding site for factor XIIa in the Apple 4 domain of coagulation factor XI. J Biol Chem 1993; 268: 3838-3844.
    PubMedGoogle Scholar
  • 10 Meijers JC, Mulvihill ER, Davie EW. et al. Apple four in human blood coagulation factor XI mediates dimer formation. Biochemistry 1992; 31: 4680-4684.
    CrossrefPubMedGoogle Scholar
  • 11 Kent WJ, Sugnet CW, Furey TS. et al. The human genome browser at UCSC. Genome Res 2002; 12: 996-1006.
    CrossrefPubMedGoogle Scholar
  • 12 Podmore A, Smith M, Savidge G. et al. Real-time quantitative PCR analysis of factor XI mRNA variants in human platelets. J Thromb Haemost 2004; 2: 1713-1719.
    CrossrefPubMedGoogle Scholar
  • 13 Asakai R, Davie EW, Chung DW. Organization of the gene for human factor XI. Biochemistry 1987; 26: 7221-7228.
    CrossrefPubMedGoogle Scholar
  • 14 Rosenthal RL, Dreskin OH, Rosenthal N. New hemophilia- like disease caused by deficiency of a third plasma thromboplastin factor. Proc Soc Exp Biol Med 1953; 82: 171-174.
    CrossrefPubMedGoogle Scholar
  • 15 Shpilberg O, Peretz H, Zivelin A. et al. One of the two common mutations causing factor XI deficiency in Ashkenazi Jews (type II) is also prevalent in Iraqi Jews, who represent the ancient gene pool of Jews. Blood 1995; 85: 429-432.
    CrossrefPubMedGoogle Scholar
  • 16 Zadra G, Asselta R, Tenchini ML. et al. Simultaneous genotyping of coagulation factor XI type II and type III mutations by multiplex real-time PCR reveals their prevalence in healthy and FXI-deficient Italians. Haematologica. 2008 in press.
    PubMedGoogle Scholar
  • 17 Mitchell M, Mountford R, Butler R. et al. Spectrum of factor XI (F11) mutations in the UK population–116 index cases and 140 mutations. Hum Mutat 2006; 27: 829.
    CrossrefPubMedGoogle Scholar
  • 18 Salomon O, Seligsohn U. New observations on factor XI deficiency. Haemophilia 2004; 10 (Suppl. 04) 184-187.
    CrossrefPubMedGoogle Scholar
  • 19 Mannucci PM, Duga S, Peyvandi F. Recessively inherited coagulation disorders. Blood 2004; 104: 1243-1252.
    CrossrefPubMedGoogle Scholar
  • 20 Kravtsov DV, Monahan PE, Gailani D. A classification system for cross-reactive material-negative factor XI deficiency. Blood 2005; 105: 4671-4673.
    CrossrefPubMedGoogle Scholar
  • 21 Kravtsov DV, Wu W, Meijers JC. et al. Dominant factor XI deficiency caused by mutations in the factor XI catalytic domain. Blood 2004; 104: 128-134.
    CrossrefPubMedGoogle Scholar
  • 22 Asakai R, Chung DW, Ratnoff OD. et al. Factor XI (plasma thromboplastin antecedent) deficiency in Ashkenazi Jews is a bleeding disorder that can result from three types of point mutations. Proc Natl Acad Sci USA 1989; 86: 7667-7671.
    CrossrefPubMedGoogle Scholar
  • 23 Peretz H, Zivelin A, Usher S. et al. A 14-bp deletion (codon 554 del AAGgtaacagagtg) at exon 14/intron N junction of the coagulation factor XI gene disrupts splicing and causes severe factor XI deficiency. Hum Mutat 1996; 8: 77-78.
    CrossrefPubMedGoogle Scholar
  • 24 Zivelin A, Bauduer F, Ducout L. et al. Factor XI deficiency in French Basques is caused predominantly by an ancestral Cys38Arg mutation in the factor XI gene. Blood 2002; 99: 2448-2454.
    CrossrefPubMedGoogle Scholar
  • 25 Quelin F, Trossaert M, Sigaud M. et al. Molecular basis of severe factor XI deficiency in seven families from the west of France. Seven novel mutations, including an ancient Q88X mutation. J Thromb Haemost 2004; 2: 71-76.
    CrossrefPubMedGoogle Scholar
  • 26 Bolton-Maggs PH, Peretz H, Butler R. et al. A common ancestral mutation (C128X) occurring in 11 non- Jewish families from the UK with factor XI deficiency. J Thromb Haemost 2004; 2: 918-924.
    CrossrefPubMedGoogle Scholar
  • 27 Reese MG, Eeckman FH, Kulp D. et al. Improved splice site detection in Genie. J Comput Biol 1997; 4: 311-323.
    CrossrefPubMedGoogle Scholar
  • 28 Rogozin IB, Milanesi L. Analysis of donor splice sites in different eukaryotic organisms. J Mol Evol 1997; 45: 50-59.
    CrossrefPubMedGoogle Scholar
  • 29 Montefusco MC, Duga S, Asselta R. et al. A novel two base pair deletion in the factor V gene associated with severe factor V deficiency. Br J Haematol 2000; 111: 1240-1246.
    CrossrefPubMedGoogle Scholar
  • 30 Dossenbach-Glaninger A, Hopmeier P. Coagulation factor XI gene analysis in three factor XI deficient Austrian patients. Eur J Haematol 2006; 76: 317-321.
    CrossrefPubMedGoogle Scholar
  • 31 Ventura C, Santos AI, Tavares A. et al. Molecular genetic analysis of factor XI deficiency: identification of five novel gene alterations and the origin of type II mutation in Portuguese families. Thromb Haemost 2000; 84: 833-840.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 32 Ars E, Serra E, Garcia J. et al. Mutations affecting mRNA splicing are the most common molecular defects in patients with neurofibromatosis type 1. Hum Mol Genet 2000; 9: 237-247.
    CrossrefPubMedGoogle Scholar
  • 33 Teraoka SN, Telatar M, Becker-Catania S. et al. Splicing defects in the ataxia-telangiectasia gene, ATM: underlying mutations and consequences. Am J Hum Genet 1999; 64: 1617-1631.
    CrossrefPubMedGoogle Scholar
  • 34 Vorechovsky I. Aberrant 3’ splice sites in human disease genes: mutation pattern, nucleotide structure and comparison of computational tools that predict their utilization. Nucleic Acids Res 2006; 34: 4630-4641.
    CrossrefPubMedGoogle Scholar
  • 35 Maquat LE, Carmichael GG. Quality control of mRNA function. Cell 2001; 104: 173-176.
    CrossrefPubMedGoogle Scholar
  • 36 Mikkola H, Syrjala M, Rasi V. et al. Deficiency in the A-subunit of coagulation factor XIII: two novel point mutations demonstrate different effects on transcript levels. Blood 1994; 84: 517-525.
    PubMedGoogle Scholar
  • 37 Soldá G, Asselta R, Ghiotto R. et al. A type II mutation (Glu117stop), induction of allele-specific mRNA degradation and factor XI deficiency. Haematologica 2005; 90: 1716-1718.
    PubMedGoogle Scholar
  • 38 Asselta R, Duga S, Spena S. et al. Congenital afibrinogenemia: mutations leading to premature termination codons in fibrinogen A alpha-chain gene are not associated with the decay of the mutant mRNAs. Blood 2001; 98: 3685-3692.
    CrossrefPubMedGoogle Scholar
  • 39 David D, Santos IM, Johnson K. et al. Analysis of the consequences of premature termination codons within factor VIII coding sequences. J Thromb Haemost 2003; 1: 139-146.
    CrossrefPubMedGoogle Scholar
  • 40 Neerman-Arbez M, Germanos-Haddad M, Tzanidakis K. et al. Expression and analysis of a split premature termination codon in FGG responsible for congenital afibrinogenemia: escape from RNA surveillance mechanisms in transfected cells. Blood 2004; 104: 3618-3623.
    CrossrefPubMedGoogle Scholar
  • 41 Gailani D, Zivelin A, Sinha D. et al. Do platelets synthesize factor XI?. J Thromb Haemost 2004; 2: 1709-1712.
    CrossrefPubMedGoogle Scholar
  • 42 Sadler J, Davie E. HemophiliaA, hemophilia B, and von Willebrand disease. Philadelphia: WB Saunders; 2001
  • 43 Bozzao C, Rimoldi V, Asselta R. et al. A novel factor XI missense mutation (Val371Ile) in the activation loop is responsible for a case of mild type II factor XI deficiency. FEBS J 2007; 274: 6128-6138.
    CrossrefPubMedGoogle Scholar
  • 44 Gailani D, Schmidt A, Sun MF. et al. A cross-reactive material positive variant of coagulation factor XI (FXIP520L) with a catalytic defect. J Thromb Haemost 2007; 5: 781-787.
    CrossrefPubMedGoogle Scholar
  • 45 Zivelin A, Ogawa T, Bulvik S. et al. Severe factor XI deficiency caused by a Gly555 to Glu mutation (factor XI-Glu555): a cross-reactive material positive variant defective in factor IX activation. J Thromb Haemost 2004; 2: 1782-1789.
    CrossrefPubMedGoogle Scholar
  • 46 Germanos-Haddad M, De Moerloose P, Boehlen F. et al. Homozygosity for a Thr575Met missense mutation in the catalytic domain associated with factor XI deficiency. Haematologica 2005; 90: 418-419.
    PubMedGoogle Scholar
  • 47 Mitchell M, Harrington P, Cutler J. et al. Eighteen unrelated patients with factor XI deficiency, four novel mutations and a 100% detection rate by denaturing high-performance liquid chromatography. Br J Haematol 2003; 121: 500-502.
    CrossrefPubMedGoogle Scholar
  • 48 O’Connell NM, Saunders RE, Lee CA. et al. Structural interpretation of 42 mutations causing factor XI deficiency using homology modeling. J Thromb Haemost 2005; 3: 127-138.
    CrossrefPubMedGoogle Scholar
  • 49 Sun MF, Baglia FA, Ho D. et al. Defective binding of factor XI-N248 to activated human platelets. Blood 2001; 98: 125-129.
    CrossrefPubMedGoogle Scholar