Identification of Eight Point Mutations in Protein S Deficiency Type I – Analysis of 15 Pedigrees

Summary

We describe molecular genetic studies of 15 patients with protein S deficiency type I (i. e. reduced total protein S antigen). All the exons of the PROS 1 gene were analyzed both by PCR and direct sequencing in all 15 probands. This analysis led to the identification of point mutations affecting eight individuals. One of these mutations (codon -25, insertion of T) has been described previously in a Dutch pedigree. The other mutations are novel and all are located in exons that code for the protein S domain that is homologous to the steroid hormone binding globulins. They include two amino acid replacements (one individual with 340 Gly → Val, and two individuals with 467 → Val Gly), and four frameshift mutations due to either one bp deletions (in codon 261 deletion of T and in codon 267 deletion of G) or insertions (in codon 565 insertion T and after codon 578 insertion of C). Studies performed in six families (totalling 43 subjects) showed cosegregation of the genetic abnormality with reduced plasma protein S levels, and provided genetic evidence for a heterozygous protein S deficiency in 25 of them. The yield of mutations in this study (53%) confirms that the percentage of protein S deficient cases in which a point mutation is found remains low.

• References

• 1 Walker FJ. The role of phospholipid in factor Va inactivation. J Biol Chem 1981; 256: 11128-11131
• 2 Comp PC, Nixon RR, Cooper MR, Esmon CT. Familial protein S deficiency is associated with recurrent thrombosis. J Clin Invest 1984; 74: 2082-2088
• 3 Suzuki K, Nishioka J, Hashimoto S. Regulation of activated protein C by thrombin-modified protein S. J Biochem (Tokyo) 1983; 94: 699-705
• 4 Koedam JA, Meijers JC, Sixma JJ, Bouma BN. Cofactor activity of protein S and protective effect of von Willebrand factor. J Clin Invest 1988; 82: 1236-1243
• 5 Heeb MJ, Mesters RM, Tans G, Rosing J, Griffin JH. Binding of protein S to factor Va associated with inhibition of prothrombinase that is independent of activated protein C. J Biol Chem 1993; 268: 2872-2877
• 6 Schwarz HP, Fischer M, Hopmeier P. Plasma protein S deficiency in familial thrombotic disease. Blood 1984; 64: 1297-1300
• 7 Broekmans AW, Bertina RM, Reinalda Poot J, Engesser L, Muller HP, Leeuw JA, Michiels JJ, Brommer EJ, Briët E. Hereditary protein S deficiency and venous thrombo-embolism. A study in three Dutch families Thromb Haemost 1985; 53: 273-277
• 8 Bertina RM. Hereditary protein S deficiency. Haemostasis 1985; 15: 241-246
• 9 Engesser L, Broekmans AW, Briët E, Brommer EJ, Bertina RM. Hereditary protein S deficiency: clinical manifestations. Ann Intern Med 1987; 106: 677-682
• 10 Tabemero MD, Tomas JF, Alberca I, Orfao A, Borrasca AL, Vicente V. Incidence and clinical characteristics of hereditary disorders associated with venous thrombosis. Am J Hematol 1991; 36: 249-254
• 11 Allaart CF, Aronson DC, Ruys T, Rosendaal FR, van Bockel JH, Bertina RM, Briët E. Hereditary protein S deficiency in young adults with arterial occlusive disease. Thromb Haemost 1990; 64: 206-210
• 12 Girolami A, Simioni P, Lazzaro AR. Heterozygous protein S deficiency: a study of a large kindred. Acta Haematol 1990; 84: 162-168
• 13 Walker FJ. Protein S and thrombotic disease. Proc Soc Exp Biol Med 1992; 200: 285-295
• 14 Ploos Van Amstel HK, Reitsma PH, van der Logt CP, Bertina RM. Intron-exon organization of the active human protein S gene PS alpha and its pseu-dogene PS beta: duplication and silencing during primate evolution. Biochemistry 1990; 29: 7853-7861
• 15 Schmidel DK, Tatso AV, Phelps LG, Tomczak JA, Long GL. Organization of the human protein S genes. Biochemistry 1990; 29: 7845-7852
• 16 Edenbrandt CM, Lindwall A, Wydro R, Stenflo J. Molecular analysis of the gene for vitamin K-dependent protein S and its pseudogene Cloning and partial gene organization. Biochemistry 1990; 29: 7861-7868
• 17 Ploos Van Amstel HK, van der Zanden AL, Bakker E, Reitsma PH, Bertina RM. Two genes homologous with human protein S cDNA are located on chromosome 3. Thromb Haemost 1987; 58: 982-987
• 18 Ploos Van Amstel HK, Reitsma PH, Bertina RM. The human protein S locus: identification of the PS alpha gene as a site of liver protein S messenger RNA synthesis. Biochem Biophys Res Commun 1988; 157: 1033-1038
• 19 Ploos Van Amstel HK, Reitsma PH, Hamulyak K, de Die Smulders CE, Mannucci PM, Bertina RM. A mutation in the protein S pseudogene is linked to protein S deficiency in a thrombophilic family. Thromb Haemost 1989; 62: 897-901
• 20 Bertina RM, Ploos Van Amstel HK, van Wijngaarden A, Coenen J, Leemhuis MP, Deutz Terlouw PP, van der Linden IK, Reitsma PH. Heerlen po-lymorphism of protein S, an immunologic polymorphism due to dimorphism of residue 460. Blood 1990; 76: 538-548
• 21 Ploos Van Amstel HK, Huisman MV, Reitsma PH, Wouter ten Cate J, Bertina RM. Partial protein S gene deletion in a family with hereditary thrombophilia. Blood 1989; 73: 479-483
• 22 Schmidel DK, Nelson RM, Broxson Jr EH, Comp PC, Marlar RA, Long GL. A 5.3-kb deletion including exon XIII of the protein S a gene occurs in two protein S-deficient families. Blood 1991; 77: 551-559
• 23 Reitsma PH, Ploos Van Amstel HK, Bertina RM. Three novel mutations in five unrelated subjects with hereditary protein S deficiency type I. J Clin Invest 1994; 93: 486-492
• 24 Gandrille S, Borgel D, Eschwege-Gufflet V, Aillaud MF, Dreyfus M, Matheron C, Gaussem P, Abgrall JF, Jude B, Sie P, Toulon P, Aiach M. Identification of 15 different candidate causal point mutations and three polymorphisms in 19 patients with protein S deficiency using a scanning method for the analysis of the protein S active gene. Blood 1995; 85: 130-139
• 25 Gómez E, Ledford MR, Pegelow CH, Reitsma PH, Bertina RM. Homozygous protein S deficiency caused by one base pair deletion that leads to a stop codon in exon III of the protein S gene. Thromb Haemost 1994; 71: 723-726
• 26 Yamazaki T, Katsumi A, Kagami K, Koyima T, Takamatsu J, Saito H. Genetic characterization of hereditary free protein S deficiency. Blood 1993; 82: 1606 (Abstract)
• 27 Hayashi T, Nishioka J, Shigekiyo T, Saito S, Suzuki K. Protein S Tokushima: Abnormal molecule with a substitution of Glu for Lys-155 in the second epidermal growth factor-like domain of protein S. Blood 1994; 83: 683-690
• 28 Jespersen J, Gram J, Bertina RM. The risk of thrombosis in hereditary protein S deficiency in a Scandinavian family. Fibrinolysis 1989; 3: 37-40
• 29 Deutz-Terlouw PP, Ballering L, van Wijngaarden A, Bertina RM. Two ELISA’s for measurement of protein S, and their use in diagnosis of protein S deficiency. Clin Chim Acta 1989; 186: 321-334
• 30 Millar SA, Dijkes DD, Polesky HF. A simple salting out procedure for extracting DNA from nucleated cells. Nucleic Acids Res 1988; 16: 1215
• 31 Saiki RK, Schaaf S, Falvona F, Mullis KB, Horn GT, Ehrlich HA, Amheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anaemia. Science 1985; 23: 1350-1354
• 32 Sanger F, Nicklen S, Coulsen AR. DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 1977; 74: 5463-5467
• 33 Chu MD, Sun J, Bird P. Cloning and sequencing of a cDNA encoding the murine vitamin K-dependent protein S. Biochim Biophys Acta 1994; 1217: 325-328
• 34 Joseph DR, Hall SH, Conti M, French FS. The gene stucture of rat androgen-binding protein: identification of potential regulatory deoxyribonucleic acid elements of a follicle-stimulating hormone regulated protein. Mol Endocrinol 1988; 2: 3-13
• 35 Gershagen S, Lundwall A, Fernlund P. Characterization of the human sex hormone binding globulin (SHBG) gene and demonstration of two transcripts both in liver and testis. Nucleic Acids Res 1989; 17: 9245-9258
• 36 Reitsma PH, Poort SR, Allaart CF, Briët E, Bertina RM. The spectrum of genetic defects in a panel of 40 Dutch families with symptomatic protein C deficiency type I: Heterogeneity and founder effects. Blood 1991; 78: 890-894
• 37 Lakich D, Kazakian HH, Antonarakis SE, Gitschier J. Inversions disrupting the factor VIII gene are a common cause of severe hemophilia A. Nature Genet 1993; 5: 236-241