Download PDF
Thromb Haemost 1994; 72(01): 065-069
DOI: 10.1055/s-0038-1648812
Original Article
Schattauer GmbH Stuttgart

Severe Homozygous Protein C Deficiency: Identification of a Splice Site Missense Mutation (184, Q → H) in Exon 7 of the Protein C Gene

J M Soria
1   The Molecular Genetics Department IRO Cancer Research Institute, Hospital Duran i Reynals, Barcelona, Spain
,
D Brito
2   Haematology Service, Hospital Regional Carlos Haya, Màlaga, Spain
,
J Barceló
2   Haematology Service, Hospital Regional Carlos Haya, Màlaga, Spain
,
J Fontcuberta
3   Haemostasis and Thrombosis Unit, Haematology Service, Hospital de Sant Pau, Barcelona, Spain
,
L Botero
2   Haematology Service, Hospital Regional Carlos Haya, Màlaga, Spain
,
J Maldonado
2   Haematology Service, Hospital Regional Carlos Haya, Màlaga, Spain
,
X Estivill
1   The Molecular Genetics Department IRO Cancer Research Institute, Hospital Duran i Reynals, Barcelona, Spain
,
N Sala
1   The Molecular Genetics Department IRO Cancer Research Institute, Hospital Duran i Reynals, Barcelona, Spain
› Author Affiliations
Further Information

Publication History

Received 22 October 1993

Accepted after resubmission 10 March 1994

Publication Date:
12 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

Single strand conformation polymorphism (SSCP) analysis of exon 7 of the protein C gene has identified a novel splice site missense mutation (184, Q → H), in a newborn child with purpura fulminans and undetectable protein C levels. The mutation, seen in the homozygous state in the child and in the heterozygous state in her mother, was characterized and found to be a G to C nucleotide substitution at the -1 position of the donor splice site of intron 7 of the protein C gene, which changes histidine 184 for glutamine (184, Q → H). According to analysis of the normal and mutated sequences, this mutation should also abolish the function of the donor splice site of intron 7 of the protein C gene. Since such a mutation is compatible with the absence of gene product in plasma and since DNA sequencing of all protein C gene exons in this patient did not reveal any other mutation, we postulate that mutation 184, Q → H results in the absence of protein C gene product in plasma, which could be the cause of the severe phenotype observed in this patient.

 

  • References

  • 1 Clouse LH, Comp PC. The regulation of haemostasis: the protein C system. New Engl J Med 1986; 314: 1298-1304
    CrossrefPubMedGoogle Scholar
  • 2 Greengard JS, Griffin JH. Protein C pathways. In: Recent Advances in Haematology Hoffbrand AV. (ed). Edinburgh, UK: Churchill Livingstone: 1988. 05 275-289
  • 3 Esmon CT. The roles of protein C and thrombomodulin in the regulation of blood coagulation. J Biol Chem 1989; 264: 4743-4746
    PubMedGoogle Scholar
  • 4 Bajzàr L, Fredenburgh JC, Nesheim M. The activated protein C-mediated enhancement of tissue-type plasminogen activator-induced fibrinolysis in a cell-free system. J Biol Chem 1990; 265: 16984-16954
    PubMedGoogle Scholar
  • 5 de Fouw NJ, van Tilburg NH, Haverkate F, Bertina RM. Activated protein C accelerates clot lysis by virtue of its anticoagulant activity. Blood Coagulation and Fibrinolysis 1993; 4: 201-210
    CrossrefPubMedGoogle Scholar
  • 6 Miletich J, Sherman L, Broze G. Absence of thrombosis in subjects with heterozygous protein C deficiency. N Engl J Med 1987; 317: 991-996
    CrossrefPubMedGoogle Scholar
  • 7 Bovill EG, Bauer KA, Dickerman JD, Callas P, West B. The clinical spectrum of heterozygous protein C deficiency in a large New England kindred. Blood 1989; 73: 712-717
    CrossrefPubMedGoogle Scholar
  • 8 Allaart CF, Poort SR, Rosendaal FR, Reitsma PH, Bertina RM, Briët E. Increased risk of venous thrombosis in carriers of hereditary protein C deficiency defect. Lancet 1993; 341: 134-138
    CrossrefPubMedGoogle Scholar
  • 9 Branson HE, Katz J, Marble R, Griffin JH. Inherited protein C deficiency and coumarin-responsive chronic relapsing purpura fulminans in a newborn infant. The Lancet 1983; 2: 1165-1168
    PubMedGoogle Scholar
  • 10 Sills RH, Marlar RA, Montgomery RR, Ganesh ND, Humbert JR. Severe homozygous protein C deficiency. J Pediatr 1984; 105: 409-413
    CrossrefPubMedGoogle Scholar
  • 11 Estellés A, García-Plaza I, Dasí A, Aznar J, Duart M, Sanz G, Pírez-Re-quejo JL, Espana F, Jimenez C, Abeldo G. Severe inherited “homozygous” protein C deficiency in a newborn infant. Thromb Haemost 1984; 52: 53-56
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Marciniak E, Wilson DH, Marlar RA. Neonatal purpura fulminans: a genetic disorder related to the absence of protein C in blood. Blood 1985; 65: 15-20
    CrossrefPubMedGoogle Scholar
  • 13 Peters C, Casella JF, Marlar RA, Montgomery RR, Zinkham WH. Homozygous protein C deficiency: observations on the nature of the molecular abnormality and the effectiveness of warfarin therapy. Pediatrics 1988; 81: 272-276
    PubMedGoogle Scholar
  • 14 Seligsohn U, Berger A, Abend M, Rubin L, Attias D, Zivelin A, Rapaport S. Homozygous protein C deficiency manifested by massive venous thrombosis in the newborn. N Engl J Med 1984; 310: 559-562
    CrossrefPubMedGoogle Scholar
  • 15 Foster DC, Yoshitake S, Davie EW. The nucleotide sequence of the gene for human protein C. Proc Natl Acad Sci USA 1985; 82: 4673-4677
    CrossrefPubMedGoogle Scholar
  • 16 Plutzky J, Hoskins JA, Long GL, Crabtree GR. Evolution and organisation of the human protein C gene. Proc Natl Acad Sci USA 1986; 83: 546-550
    CrossrefPubMedGoogle Scholar
  • 17 Rocchi M, Roncuzzi L, Santamaria R, Archidiacono N, Dente L, Romeo G. Mapping through somatic cell hybrids and cDNA probes of protein C to chromosome 2, factor X to chromosome 13 and α1-acid glycoprotein to chromosome 9. Hum Genet 1986; 74: 30-33
    PubMedGoogle Scholar
  • 18 Reitsma PH, Poort SR, Bemardi F, Gandrille S, Long GL, Sala N, Cooper DN. Protein C deficiency: A database of mutations. Thromb Haemost 1993; 69: 77-84
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Matsuda M, Sugo T, Sakata Y, Murayama H, Mimuro J, Tanabe S, Yoshitake S. A thrombotic state due to an abnormal protein C. New Engl J Med 1988; 319: 1265-1268
    CrossrefPubMedGoogle Scholar
  • 20 Grundy CB, Melissari E, Undo V, Scully MF, Kakkar VV, Cooper DN. Late-onset homozygous protein C deficiency. Lancet 1991; 338: 575-576
    PubMedGoogle Scholar
  • 21 Conard J, Horellou MH, van Dreden P, Samama M, Reitsma PH, Poort S, Bertina RM. Homozygous protein C deficiency with late onset and recurrent coumarin-induced skin necrosis. Lancet 1992; 339: 743-744
    CrossrefPubMedGoogle Scholar
  • 22 Grundy CB, Chisholm M, Kakkar VV, Cooper DN. A novel homozygous missense mutation in the protein C (PROC) gene causing recurrent venous thrombosis. Hum Gent 1992; 89: 683-684
    PubMedGoogle Scholar
  • 23 Sugahara Y, Miura O, Yuen P, Aoki N. Protein C deficiency Hong Kong 1 and 2: Hereditary protein C deficiency caused by two mutant alleles, a 5-nucleotide deletion and a missense mutation. Blood 1992; 80: 126-133
    PubMedGoogle Scholar
  • 24 Ido M, Ohiwa M, Hayashi T, Nishioka J, Hatada T, Watanabe Y, Wada H, Shirakawa S, Suzuki K. A compound heterozygous protein C deficiency with a single nucleotide G deletion encoding Gly-381 and an aminoacid substitution of Lys for Gla-26. Thromb Haemost 1993; 70: 636-641
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucl Acids Res 1988; 16: 1215
    CrossrefPubMedGoogle Scholar
  • 26 Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K. Current Protocols in Molecular Biology. Current Protocols, John Wiley and Sons, Inc 1993 unit 2.9A.
    PubMedGoogle Scholar
  • 27 Berg LP, Wieland K, Millar DS, Schlösser M, Wagner M, Kakkar VV, Reiss J, Cooper DN. Detection of a novel point mutation causing haemophilia A by PCR/direct sequencing of ectopically transcribed factor VIII mRNA. Human Genetics 1990; 85: 655-658
    PubMedGoogle Scholar
  • 28 Feinberg AP, Vogelstein B. A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-13
    CrossrefPubMedGoogle Scholar
  • 29 Reitsma PH, Poort SR, Allaart CF, Briët E, Bertina R. The spectrum of genetic defects in a panel of 40 Dutch families with symptomatic protein C deficiency type 1: Heterogeneity and founder effects. Blood 1991; 78: 890-894
    PubMedGoogle Scholar
  • 30 Soria JM, Fontcuberta J, Borrell M, Estivill X, Sala N. Protein C deficiency: Identification of a novel two-base pair insertion and two point mutations in exon 7 of the protein C gene in Spanish families. Human Mutation 1992; 1: 428-431
    CrossrefPubMedGoogle Scholar
  • 31 Soria JM, Fontcuberta J, Chillón M, Borrell M, Estivill X, Sala N. Acceptor splice site mutation in the invariant AG of intron 5 of the protein C gene, causing type I protein C deficiency. Hum Genet 1993; 92: 506-508
    CrossrefPubMedGoogle Scholar
  • 32 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: 7155-7174
    CrossrefPubMedGoogle Scholar
  • 33 Senepathy P, Shapiro MB, Harris NL. Splice junctions, branch point sites and exons: sequence statistics, identification and applications to genome project. Methods Enzymol 1990; 183: 252-278
    PubMedGoogle Scholar
  • 34 Morral N, Nunes V, Casals T, Cobos N, Asensio O, Dapena J, Estivill X. Uniparental inheritance of microsatellite alleles of the cystic fibrosis gene (CFTR): Identification of a 50 kb deletion. Human Molecular Genetics 1993; 2: 677-681
    CrossrefPubMedGoogle Scholar
  • 35 Soria JM, Ibáñez I, Fontcuberta J, Borrell M, Estivill X, Sala N. Apliciación del polimorfismo MI, reconocido por la enzima MspI, al estudio del déficit congénito de proteýna C.. Med Clin (Bare) 1992; 99: 649-652
    PubMedGoogle Scholar
  • 36 Koenhen E, Bertina RM, Reitsma PH. MspI RFLP in intron 8 of the protein C gene. Nucl Acad Res 1989; 17: 8401
    PubMedGoogle Scholar
  • 37 Marlar RA, Montgomery RR, Broekmans AW. et al. Diagnosis and treatment of homozygous protein C deficiency. J Pediatr 1989; 114: 528-534
    CrossrefPubMedGoogle Scholar
  • 38 Marlar RA, Sills RH, Groncy PK, Montgomery RR, Madden RM. Protein C survival during replacement therapy in homozygous protein C deficiency. Am J Hematol 1992; 41: 24-31
    CrossrefPubMedGoogle Scholar
  • 39 Krawczak M, Reiss J, Cooper DN. The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: causes and consequences. Hum Genet 1992; 90: 41-54
    PubMedGoogle Scholar
  • 40 Lind B, van Solinge WW, Schwartz M, Thorsen S. Splice site mutation in the human protein C gene associated with venous thrombosis: demonstration of exon skipping by ectopic transcript analysis. Blood 1993; 82: 2423-2432
    PubMedGoogle Scholar