Severe Factor VII Deficiency Caused by a Novel Mutation His³⁴⁸ to Gln in the Catalytic Domain

Summary

Factor VII is a vitamin K-dependent zymogen that plays a key role in the initiation of the extrinsic pathway. A severe factor VII deficiency was identified in a 45-year old male whose plasma factor VII antigen was less than 60 ng/ml and expressed 5.2% of normal factor VII activity. DNA sequence analysis of the patient’s factor VII gene showed a thymidine to guanine transversion at nucleotide 10968 in exon VIII that results in a novel amino acid substitution of His³⁴⁸ to Gln. The patient was homozygous for this mutation, whereas some of his family members were heterozygous. Both wild type and mutant factor VII were transiently expressed in COS-1 cells. The level of secreted mutant factor VII antigen was only 11.0% of the level of wild type factor VII. In CHO cells stably transfected with the mutant factor VII, only 37.3% of the total labeled FVII was secreted into the conditioned media and the remainder was retained inside the cells. These data suggest this mutation leads to factor VII deficiency due to the impaired secretion of the molecule.

Key words

Factor VII - catalytic domain - type I deficiency - impaired secretion

³ Present address: Dr. A. Katsumi, Howard Hughes Medical Institute Research Laboratories, Washington University School of Medicine, 660 South Euclid Avenue, Box 8022, St. Louis, MO 63110, USA.

• References

• 1 Davie EW, Fujikawa K, Kisiel W. The coagulation cascade: initiation, maintenance, and regulation. Biochemistry 1991; 30: 10363-70.
• 2 Fair DS. Quantitation of factor VII in the plasma of normal and warfarintreated individuals by radioimmunoassay. Blood 1983; 62: 784-91.
• 3 Cooper DN, Millar DS, Wacey A, Banner DW, Tuddenham EGD. Inherited factor VII deficiency: Molecular genetics and pathophysiology. Thromb Haemost 1997; 78: 151-60.
• 4 Kamiya T, Sugihara T, Ogata K, Saito H, Suzuki K, Nishioka J, Hashimoto S, Yamagata K. Inherited deficiency of protein S in a Japanese family with recurrent venous thrombosis: a study of three generations. Blood 1986; 67: 406-10.
• 5 Matsushita T, Kojima T, Emi N, Takahashi I, Saito H. Impaired human tissue factor-mediated activity in blood clotting factor VII Nagoya (Arg304---Trp) Evidence that a region in the catalytic domain of factor VII is important for the association with tissue factor. J Biol Chem 1994; 269: 7355-63.
• 6 Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487-91.
• 7 Matsushita T, Tanimoto M, Yamamoto K, Sugiura I, Hamaguchi M, Takamatsu J, Kamiya T, Saito H. DNA sequence analysis of three inhibitorpositive hemophilia B patients without gross gene deletion: identification of four novel mutations in factor IX gene. J Lab Clin Med 1990; 116: 492-7.
• 8 O’Hara PJ, Grant FJ, Haldeman BA, Gray CL, Insley MY, Hagen FS, Murray MJ. Nucleotide sequence of the gene coding for human factor VII, a vitamin K-dependent protein participating in blood coagulation. Proc Natl Acad Sci USA 1987; 84: 5158-62.
• 9 Di Castelnuovo A, D’Orazio A, Amore C, Falanga A, Kluft C, Donati MB, Iacoviello L. Genetic modulation of coagulation factor VII plasma levels: contribution of different polymorphisms and gender-related effects. Thromb Haemost 1998; 80: 592-7.
• 10 Katsumi A, Kojima T, Senda T, Yamazaki T, Tsukamoto H, Sugiura I, Kobayashi S, Miyata T, Umeyama H, Saito H. The Carboxyl-terminal region of protein C is essential for its secretion. Blood 1998; 91: 3784-91.
• 11 Kaufman RJ, Davies MV, Wasley LC, Michnick D. Improved vectors for stable expression of foreign genes in mammalian cells by use of the untranslated leader sequence from EMC virus. Nucleic Acids Res 1991; 19: 4485-90.
• 12 Katsumi A, Senda T, Yamashita Y, Yamazaki T, Hamaguchi M, Kojima T, Kobayashi S, Saito H. Protein C Nagoya, an elongated mutant of protein C, is retained within the endoplasmic reticulum and is associated with GRP78 and GRP94. Blood 1996; 87: 4164-75.
• 13 Green F, Kelleher C, Wilkes H, Temple A, Meade T, Humphries S. A common genetic polymorphism associated with lower coagulation factor VII levels in healthy individuals. Arterioscler Thromb 1991; 11: 540-6.
• 14 Marchetti G, Patracchini P, Papacchini M, Ferrati M, Bernardi F. A polymorphism in the 5’ region of coagulation factor VII gene (F7) caused by an inserted decanucleotide. Hum Genet 1993; 90: 575-6.
• 15 Yamazaki T, Katsumi A, Kagami K, Okamoto Y, Sugiura I, Hamaguchi M, Kojima T, Takamatsu J, Saito H. Molecular basis of a hereditary type I protein S deficiency caused by a substitution of Cys for Arg474. Blood 1996; 87: 4643-50.
• 16 Tokunaga F, Tsukamoto T, Koide T. Cellular basis for protein C deficiency caused by a single amino acid substitution at Arg15 in the gamma-carboxyglutamic acid domain. J Biochem (Tokyo) 1996; 120: 360-8.
• 17 Hunault M, Arbini AA, Carew JA, Peyvandi F, Bauer KA. Characterization of two naturally occurring mutations in the second epidermal growth factor-like domain of factor VII. Blood 1999; 93: 1237-44.
• 18 Greenberg CS, Orthner CL. Blood Coagulation and Fibrinolysis. In: Wintrobe’s Clinical Hematology. Lee GR, Foerster J, Lukens J, Paraskevas F, Greer JP, Rodgers GM. eds. Baltimore: Williams &: Wilkins; 1998: 684-764.
• 19 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: 267-70.
• 20 Bottema CD, Ketterling RP, Yoon HS, Sommer SS. The pattern of factor IX germ-line mutation in Asians is similar to that of Caucasians. Am J Hum Genet 1990; 47: 835-41.
• 21 Odom MW, Leone G, De Stefano V, Montiel MM, Boland EJ, Anderson J, Jagadeeswaran P. Five novel point mutations: two causing haemophilia B and three causing factor X deficiency. Mol Cell Probes 1994; 08: 63-5.
• 22 Banner DW, D’Arcy A, Chene C, Winkler FK, Guha A, Konigsberg WH, Nemerson Y, Kirchhofer D. The crystal structure of the complex of blood coagulation factor VIIa with soluble tissue factor. Nature 1996; 380: 41-6.