Characterization of a Mouse Factor IX cDNA and Developmental Regulation of the Factor IX Gene Expression in Liver^[*]

Summary

A mouse factor IX cDNA was isolated and characterrzed. The cDNA was 1,837 bp in length and contained the coding region as well as short 5’ and 3’ untranslated sequences. Northern blot analysis of liver RNA showed two mRNA species of 3.2 kb (major) and 2.2 kb (minor) for the mouse factor IX. An antisgnse RNA probe prepared from the mouse cDNA was employed to determine the steady state level of factor IX mRNA in mouse liver at various developmental stages. The factor IX mRNA level was very low (2–5% of the adult level) during the gestational period until day –3 (gestational day 17) followed by a rapid increase at day –2 through birth. This phase of rapid increase was followed by a gradual increase before it reached the adult level at around 20 to 24 days. At birth, the factor IX mRNA level was found to be at about 43% of that of the adult. The rnRNA levels in mouse liver agreed well with the plasma factor IX activity levels. These results indicate that reduced factor IX activity in newborns is due to the low levels of factor IX mRNA available for translation.

^* This work was supported in part by research grants from the National Institutes of Health (HL38644 and HL39099) and a research grant from the Hemophilia Foundation of Michigan to K. K.

• References

• 1 Kurachi K, Chen S-H. Human genes for factor IX and other vitamin K dependent blood proteins. In: The New Dimension of Warfarin Prophylaxis Wesler S, Becker CG, Nemerson Y. (eds) Plenum Publishing Corp; New York: 1987: 67-81
• 2 Hedner U, Davie EW. Introduction to hemostasis and the vitamin Independent coagulation factor. In: The Metabolic Basis of Inherited Diseases, Vol. II. Sixth ed Scriner CR, Beaudet AL, Sly WS, Valle D. (eds) McGraw Hill; New York: 1989: 2107-2134
• 3 Salier JP, Hirosawa S, Kurachi K. Functional characterization of the 5’ regulatory region of human factor IX gene. J Biol Chem 1990; 265: 7062-7068
• 4 Kurachi K, Chen SH. Molecular genetic analysis of congenital disease: Disorders of human blood coagulation factors. In: Hematologic Diseases in Maternal-Fetal Medicine Bern MM, Frigoletto F. (eds) Alan R. Liss, Inc; New York: 1990. in press
• 5 Kurachi K, Davie EW. Isolation and characterization of a cDNA coding for human factor IX gene. Proc Natl Acad Sci USA 1982; 79: 6461-6464
• 6 Yoshitake S, Schack BG, Foster DC, Davie EW, Kurachi K. Nucleotide sequence of the gene for human factor IX. Biochemistry 1985; 24: 3736-3750
• 7 Anson DS, Choo KH, Rees D JG, Giannelli F, Gould K, Huddelston JA, Brownlee GG. The gene structure of human antihemophilic factor IX. EMBO J 1984; 3: 1053-1060
• 8 Jaye M, de la SalleH, Schamber F, Balland A, Kohli V, Findeli A, Tolstoshev P, Lecocq JP. Isolation of a human anti-hemophilic factor IX cDNA clone using a unique 52-base synthetic oligonucleotide probe deduced from the amino acid sequence of bovine factor IX. Nucl Acid Res 1983; 11: 2325-2335
• 9 Hirosawa S, Fahner JB, Salier JP, Wu CT, Lovrien EW, Kurachi K. Structural and functional basis of the developmental regulation of human coagulation factor IX gene: Factor IX-Leyden. Proc Natl Acad Sci USA 1990; 87: 4421-4425
• 10 Terwiel JP, Veltkamp JJ, Bertina RM, Muller HP. Coagulation factors in the human fetus of about 20 weeks of gestational age. Br J Haematol 1980; 45: 641-650
• 11 Schettini F, Mattia DD, Altomare M, Montagna O. Postnatal development of factor IX. Acta Paed Scand 1980; 69: 53-58
• 12 Thompson AR. Factor IX and prothrombin in amniotic fluid and fetal plasma: constraints on prenatal diagnosis of hemophilia B and evidence of proteolysis. Blood 1984; 64: 867-874
• 13 Forestier F, Daffos F, Rainaut M, Sole Y, Amiral J. Vitamin K dependent proteins in fetal hemostasis at mid trimester of pregnancy. Thromb Haemostas 1985; 53: 401-403
• 14 Andrew M, Paes B, Milner R, Johnston M, Mitchel L, Tollefsen DM, Powers P. Development of the human coagulation system in the full-term infant. Blood 1987; 70: 165-172
• 15 Andrew M, Paes B, Milner R, Johnston M, Mitchel L, Tollefsen DM, Castle V, Powers P. Development of the human coagulation system in the healthy premature infant. Blood 1988; 72: 1651-1657
• 16 Kisker CT, Robillard JE, Clarke WR. Development of blood coagulation – a fetal lamb model. Paediatr Res 1981; 15: 1045-1050
• 17 Kisker CT, Perlman S, Bohlken D, Wicklund B. Measurement of prothrombin mRNA during gestation and early neonatal development. J Lab Clin Med 1988; 112: 407-412
• 18 Lane PA, Hathaway WE. Vitamin K in infancy. J Pediat 1985; 106: 351-359
• 19 Malia RG, Preston FE, Mitchell VE. Evidence against vitamin K deficiency in normal neonates. Thromb Haemostas 1980; 44s: 159-160
• 20 Bloch CA, Rothberg AD, Bradlow BA. Mother-infant prothrombin precursor status at birth. J Paediat Gastoenterol Nutr 1984; 3: 101-103
• 21 Benton WD, Davis RW. Screening λgt recombinant clones by hybridization to single plaques in situ. Science 1977; 196: 180-182
• 22 Ausubel FM, Brent R, Kingston RE, Moore DD, Smith JA, Seidman JG, Struhl K. In: Current Protocols in Molecular Biology John Wiley and Sons; New York: 1987
• 23 Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 1977; 74: 5463-5467
• 24 Samuelson LC, Keller PR, Darlington GJ, Meisler MH. Glucocorticoid and developmental regulation of amylase mRNA in mouse liver cells. Mol Cell Biol 1988; 8: 3857-3863
• 25 Sambrook J, Fritsch EF, Maniatis T. In: Molecular Cloning, A Laboratory Manual. 2nd ed Cold Spring Harbor Laboratory Press; New York: 1989: 7.37-7.52
• 26 Evans JP, Watzke HH, Ware JL, Stafford DW, High KA. Molecular cloning of a cDNA encoding canine factor IX. Blood 1989; 74: 207-212
• 27 Sarker G, Koeberl DD, Sommer SS. Direct sequencing of the activation peptide and catalytic domain of the factor IX gene in six species. Genomics 1990; 6: 133-143
• 28 Wu SM, Stafford DW, Ware J. Deduced amino acid sequence of mouse blood coagulation factor IX. Gene 1990; 86: 275-278
• 29 Katayama K, Ericsson LH, Enfield DL, Thompson CAB, Walsh KA, Neurath H, Davie EW, Titani K. Comparison of amino acid sequence of bovine coagulation factor IX (Christmas factor) with that of other vitamin-K dependent plasma proteins. Proc Natl Acad Sci USA 1979; 76: 4990-4994
• 30 Hassan HJ, Leonardi A, Chelucci C, Guerriero R, Mannucci PM, Peschle C. Expression in ontogenesis of human blood coagulation factors. Thromb Haemostas 1987; 58: 490
• 31 Thompson AR. Factor IX antigen by radioimmunoassay. Abnormal factor IX protein in patients on warfarin and with hemophilia B. J Clin Invest 1977; 59: 900-910