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Thromb Haemost 2000; 84(02): 307-311
DOI: 10.1055/s-0037-1614012
Review Article
Schattauer GmbH

A Novel Point Mutation of the Splicing Donor Site in the Intron 2 of the Plasmin Inhibitor Gene

Haruhiko Yoshinaga
1   From the First Department of Internal Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan
,
Shinsaku Hirosawa
1   From the First Department of Internal Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan
,
Dong Hui Chung
1   From the First Department of Internal Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan
,
Nobuyuki Miyasaka
1   From the First Department of Internal Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan
,
Nobuo Aoki
1   From the First Department of Internal Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan
,
Remi Favier
2   Laboratory of Hematology and Hemostasis, Hospital Armand Trousseau, Paris, France
› Author AffiliationsThis study was supported in part by a research grant to R. F. from the French Society of Pediatric Pathology and grants to S. H. for intractable diseases from the Ministry of Health and Welfare of Japan.
Further Information

Publication History

Received 07 December 1999

Accepted after revision 16 March 2000

Publication Date:
14 December 2017 (online)

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Summary

Plasmin inhibitor (PI) is a major physiological inhibitor of plasminmediated fibrinolysis; hence, its deficiency results in a severe haemorrhagic diathesis. We analyzed the PI gene of a French boy apparently homozygous for PI deficiency and his heterozygous parents. Both alleles of the homozygous patient had a novel G to A transition at the consensus splicing donor site in the intron 2 of the PI gene. In an expression assay using the heterologous cells transfected with the mutant PI expression vector, 3 types of aberrant transcripts using a cryptic splicing donor site within the intron 2 were detected. All of these mRNAs had a stop codon upstream of the cryptic splicing site and encode only 25 amino acids, comprising the first 21 amino acids of the signal peptide (27 amino acids) plus 4 new amino acids. This mutant was designated as PI-Paris-Trousseau.

Keywords

Plasmin inhibitor - mutation - fibrinolysis - splicing donor site
 

  • References

  • 1 Moroi M, Aoki N. Isolation and characterization of α2-plasmin inhibitor from human plasma. A novel proteinase inhibitor which inhibits activatorinduced clot lysis. J Biol Chem 1976; 251: 5956-65.
    PubMedGoogle Scholar
  • 2 Aoki N, Moroi M, Matsuda M, Tachiya K. The behavior of α2-plasmin inhibitor in fibrinolytic states. J Clin Invest 1977; 60: 361-9.
    CrossrefPubMedGoogle Scholar
  • 3 Aoki N, Harpel PC. Inhibitors of the fibrinolytic enzyme system. Semin Thromb Hemost 1984; 10: 24-41.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Wiman B, Collen D. On the kinetics of the reaction between human antiplasmin and plasmin. Eur J Biochem 1978; 84: 573-8.
    CrossrefPubMedGoogle Scholar
  • 5 Aoki N, Moroi M, Tachiya K. Effects of α2-plasmin inhibitor on fibrin clot lysis. Its comparison with α2-macroglobulin. Thromb Haemost 1978; 39: 22-31.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Sakata Y, Aoki N. Cross-linking of α2-plasmin inhibitor to fibrin by fibrinstabilizing factor. J Clin Invest 1980; 65: 290-7.
    CrossrefPubMedGoogle Scholar
  • 7 Sakata Y, Aoki N. Significance of cross-linking of α2-plasmin inhibitor to fibrin in inhibition of fibrinolysis and in hemostasis. J Clin Invest 1982; 69: 536-42.
    CrossrefPubMedGoogle Scholar
  • 8 Jansen JW, Haverkate F, Koopman J, Nieuwenhuis HK, Kluft C, Boschman TA. Influence of factor XIIIa activity on human whole blood clot lysis in vitro. Thromb Haemost 1987; 57: 171-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Bangert K, Johnsen AH, Christensen U, Thorsen S. Different N-terminal forms of α2-plasmin inhibitor in human plasma. Biochem J 1993; 291: 623-5.
    CrossrefPubMedGoogle Scholar
  • 10 Koyama T, Koike Y, Toyota S, Miyagi F, Suzuki N, Aoki N. Different NH2-terminal form with 12 additional residues of α2-plasmin inhibitor from human plasma and culture media of Hep G2 cells. Biochem Biophys Res Commun 1994; 200: 417-22.
    CrossrefPubMedGoogle Scholar
  • 11 Sumi Y, Ichikawa Y, Nakamura Y, Miura O, Aoki N. Expression and characterization of the pro α2-plasmin inhibitor. J Biochem 1989; 106: 703-7.
    CrossrefPubMedGoogle Scholar
  • 12 Hirosawa S, Nakamura Y, Miura O, Sumi Y, Aoki N. Organization of the human α2-plasmin inhibitor gene. Proc Natl Acad Sci USA 1988; 85: 6836-40.
    CrossrefPubMedGoogle Scholar
  • 13 Kato A, Hirosawa S, Toyota S, Nakamura Y, Nishi H, Kimura A, Sasazuki T, Aoki N. Localization of the human α2-plasmin inhibitor gene (PLI) to 17p13. Cytogenet Cell Genet 1993; 62: 190-1.
    CrossrefPubMedGoogle Scholar
  • 14 Aoki N, Saito H, Kamiya T, Koie K, Sakata Y, Kobakura M. Congenital deficiency of α2-plasmin inhibitor associated with severe hemorrhagic tendency. J Clin Invest 1979; 63: 877-84.
    CrossrefPubMedGoogle Scholar
  • 15 Aoki N, Sakata Y, Matsuda M, Tateno K. Fibrinolytic states in a patient with congenital deficiency of α2-plasmin inhibitor. Blood 1980; 55: 483-8.
    PubMedGoogle Scholar
  • 16 Saito H. α2-plasmin inhibitor and its deficiency states. J Lab Clin Med 1988; 112: 671-8.
    PubMedGoogle Scholar
  • 17 Aoki N. Hemostasis associated with abnormalities of fibrinolysis. Blood Rev 1989; 03: 11-7.
    CrossrefPubMedGoogle Scholar
  • 18 Kluft C, Nieuwenhuis HK, Rijken DC, Groeneveld E, Hijngaards G, van Berke W, Dooijewaards G, Sixma JJ. α2-Antiplasmin Enschede: dysfunctional α2-antiplasmin molecule associated with an autosomal recessive hemorrhagic disorder. J Clin Invest 1987; 80: 1391-400.
    CrossrefPubMedGoogle Scholar
  • 19 Holmes WE, Lijnen HR, Nelles L, Kluft C, Nieuwenhuis HK, Rijken DC, Collen D. α2-antiplasmin Enschede: alanine insertion and abolition of plasmin inhibitory activity. Science 1987; 238: 209-11.
    CrossrefPubMedGoogle Scholar
  • 20 Miura O, Hirosawa S, Kato A, Aoki N. Molecular basis for congenital deficiency of α2-plasmin inhibitor. A frameshift mutation leading to elongation of the deduced amino acid sequence. J Clin Invest 1989; 83: 1598-604.
    CrossrefPubMedGoogle Scholar
  • 21 Miura O, Sugahara Y, Aoki N. Hereditary α2-plasmin inhibitor deficiency caused by a transportdeficient mutation (α2-PI-Okinawa). Deletion of Glu137 by a trinucleotide deletion blocks intracellular transport. J Biol Chem 1989; 264: 18213-9.
    PubMedGoogle Scholar
  • 22 Miura O, Aoki N. Impaired secretion of mutant α2-plasmin inhibitor (α2PINara) from COS-7 and HepG2 cells: molecular and cellular basis for hereditary deficiency of α2-plasmin inhibitor. Blood 1990; 75: 1092-6.
    PubMedGoogle Scholar
  • 23 Toyota S, Hirosawa S, Aoki N. Secretion of α2-plasmin inhibitor is impaired by amino acid deletion in a small region of the molecule. J Biochem (Tokyo) 1994; 115: 293-7.
    CrossrefPubMedGoogle Scholar
  • 24 Paqueron X, Favier R, Richard P, Maillet J, Murat I. Severe postadenoidectomy bleeding revealing congenital α2 antiplasmin deficiency in a child. Anesth Analg 1997; 84: 1147-9.
    CrossrefPubMedGoogle Scholar
  • 25 Devaussuzenet VMP, Ducou-le-Pointe HA, Doco AM, Mary PM, Montagne JR, Favier R. A case of intramedullary haematoma associated with congenital α2-plasmin inhibitor deficiency. Pediatr Radiol 1998; 28: 978-80.
    CrossrefPubMedGoogle Scholar
  • 26 Giannelli F, Green PM, Sommer SS, Poon M, Ludwig M, Schwaab R, Reitsma PH, Goossens M, Yoshioka A, Figueiredo MS, Brownlee GG. Haemophilia B: database of point mutations and short additions and deletions – eighth edition. Nucleic Acids Res 1998; 26: 265-8.
    CrossrefPubMedGoogle Scholar
  • 27 Reitsma PH, Poort SR, Bernardi F, Gandrille S, Long GL, Sala N, Cooper DN. Protein C deficiency: a database of mutations. For the Protein C & S Subcommittee of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis. Thromb Haemost 1993; 69: 77-84.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Krawczak M, Reiss J, Cooper DN. The mutational spectrum of single basepair substitutions in mRNA splice junctions of human genes: causes and consequences. Hum Genet 1992; 90: 41-54.
    PubMedGoogle Scholar
  • 29 Treisman R, Proudfoot NJ, Shander M, Maniatis T. A single-base change at a splice site in a β0-thalassemic gene causes abnormal RNA splicing. Cell 1982; 29: 903-11.
    CrossrefPubMedGoogle Scholar
  • 30 Berg LP, Grundy CB, Thomas F, Millar DS, Green PJ, Slomski R, Reiss J, Kakkar W, Cooper DN. De novo splice site mutation in the antithrombin III (AT3) gene causing recurrent venous thrombosis: demonstration of exon skipping by ectopic transcript analysis. Genomics 1992; 13: 1359-61.
    CrossrefPubMedGoogle Scholar
  • 31 Shapiro MB, Senapathy P. RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res 1987; 15: 7155-74.
    CrossrefPubMedGoogle Scholar
  • 32 Robberson BL, Cote GJ, Berget SM. Exon definition may facilitate splice site selection in RNAs with multiple exons. Mol Cell Biol 1990; 10: 84-94.
    CrossrefPubMedGoogle Scholar
  • 33 Chung DH, Ohashi K, Watanabe M, Miyasaka N, Hirosawa S. Mannose trimming targets mutant α2-plasmin inhibitor for degradation by the proteasome. J Biol Chem 2000; 275: 4981-7.
    CrossrefPubMedGoogle Scholar