Detection of Ten New Mutations by Screening the Gene Encoding Factor IX of Danish Hemophilia B Patients

Summary

Hemophilia B is caused by a wide range of mutations. In order to characterize the mutations among patients in Denmark, we have systematically screened the entire coding region, the promoter region and exon flanking sequences of the gene encoding factor IX using single strand conformation and heteroduplex analyses. Patients from 32 different families were examined, and point mutations (23 different) were found in all of them. Ten of the mutations have not been reported by others; they include a splice site mutation, a single base pair deletion, and missense mutations. Notably, the study contains a female patient and a previously described Leyden mutation. In ten families with sporadic cases of hemophilia B, all 10 mothers were found to be carriers. The origin of two of these mutations was established.

• References

• 1 Yoshitake S, Schach BG, Foster DC, Davie EW, Kurachi K. Nucleotide sequence of the gene for Human Factor IX (Antihemophilic Factor B). Biochemistry 1985; 24: 3736-3750
• 2 Roberts HR. Molecular biology of hemophilia B. Thromb Haemost 1993; 70: 1-9
• 3 Bottema CD K, Ketterling RP, Ii S, Yoon HS, Phillips III JA, Sommer SS. Missense mutations and evolutionary conservation of amino acids: evi-dence that many of the amino acids in factor IX function as “spacer” elements. Am J Hum Genet 1991; 49: 820-838
• 4 Anson DS, Choo KH, Rees DJ G, Giannelli F, Gould K, Huddleston JA, Brownlee GG. The gene structure of human anti-haemophilic factor I X. EMBOJ 1984; 3: 1053-1060
• 5 Green PM, Bentley DR, Mibashan RS, Nilsson IM, Giannelli F. Molecular pathology of haemophilia B. EMBO J 1989; 8: 1067-1072
• 6 Orita M, Suzuki Y, Sekiya T, Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 1989; 5: 874-879
• 7 White MB, Carvalho M, Derse D, O’Brien SJ, Dean M. Detecting single base substitutions as heteroduplex polymorphisms. Genomics 1992; 12: 301-306
• 8 Gyllensten UB, Erlich HA. Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sei USA 1988; 85: 7652-7656
• 9 Reitsma PH, Bertina RM, Ploos van Amstel JK, Riemens A, Briët E. The putative factor IX gene promoter in hemophilia B Leyden. Blood 1988; 72: 1074-1076
• 10 McGraw RA, Davis LM, Noyes CM, Lundblad RL, Roberts HR, Graham JB, Stafford DW. Evidence for a prevalent dimorphism in the activation peptide of human coagulation factor IX. Proc Natl Acad Sci USA 1985; 82: 2847-2851
• 11 Giannelli F, Green PM, Sommer SS, Lillicrap DP, Ludwig M, Schwaab R, Reitsma PH, Goossens M, Yoshioka A, Brownlee GG. Haemophilia B: database of point mutations and short additions and deletions, fifth edition, 1994. Nucleic Acids Res 1994; 22: 3534-3546
• 12 Bottema CD K, Bottema MJ, Ketterling RP, Yoon HS, Janco RL, Phillips III JA, Sommer SS. Why does the human factor IX gene have a G + C content of 40%. Am J Hum Genet 1991; 49: 839-850
• 13 Zhang L, Jhingan A, Castellino FJ. Role of individual γ-carboxyglutamic acid residues of activated human protein C in defining its in vitro anticoagulant activity. Blood 1992; 80: 942-952
• 14 Nielsen LR, Schwartz M, Scheibel E. Screening for mutations in the gene encoding factor I X. J Inherit Metab Dis 1992; 15: 339-341
• 15 Noyes CM, Griffith MJ, Roberts HR, Lundblad RL. Identification of the molecular defect in factor IX_{Chapel Hill}: Substitution of histidine for arginine at position 145. Proc Natl Acad Sci USA 1983; 80: 4200-4202
• 16 Liddell MB, Peake IR, Taylor SA M, Lillicrap DP, Giddings JC, Bloom AL. Factor IX Cardiff: a variant factor IX protein that shows abnormal activation is caused by an arginine to cysteine substitution at position 145. Br J Haematol 1989; 72: 556-560
• 17 Miyata T, Sakai T, Sugimoto M, Naka H, Yamamoto K, Yoshioka A, Fukui H, Mitsui K, Kamiya K, Umeyama H, Iwanaga S. Factor IX Amaga-saki: a new mutation in the catalytic domain resulting in the loss of both coagulant and esterase activities. Biochemistry 1991; 30: 11286-11291
• 18 Green PM, Montandon AJ, Ljung R, Nilsson IM, Giannelli F. Haplotype analysis of identical factor IX mutants using PCR. Thromb Haemost 1992; 67: 66-69
• 19 Efstratiadis A, Posakony JW, Maniatis T, Lawn RM, O’Connell C, Spritz RA, DeRiel JK, Forget BG, Weissmann SM, Slightom JL, Blechl AE, Smithies O, Baralle FE, Shoulders CC, Proudfoot NJ. The structure and evolution of the human β-globin gene family. Cell 1980; 21: 653-668
• 20 Ghanem N, Costes B, Matin J, Vidaud M, Rothschild C, Foyer-Gazengel C, Goossens M. Twenty-four novel hemophilia B mutations revealed by rapid scanning of the whole factor IX gene in a French population sample. Eur J Hum Genet 1993; 1: 144-155
• 21 Tartary M, Vidaud D, Piao Y, Costa JM, Bahnak BR, Fressinaud E, Con-gard B, Laurian Y, Meyer D, Lavergne JM, Vidaud M. Detection of a molecular defect in 40 of 44 patients with haemophilia B by PCR and denaturing gradient gel electrophoresis. Br J Haematol 1993; 84: 662-669
• 22 Green PM, Montandon AJ, Ljung R, Bentley DR, Nilsson IM, Kling S, Giannelli F. Haemophilia B mutations in a complete Swedish population sample: a test of new strategy for the genetic counselling of diseases with high mutational heterogeneity. Br J Haematol 1991; 78: 390-397
• 23 Lin SW, Shen MC. Genetic basis and carrier detection of hemophilia B of Chinese origin. Thromb Haemost 1993; 69: 247-252