New Splicing Mutations in the Human Factor XIIIA Gene, Each Producing Multiple Mutant Transcripts of Varying Abundance

Summary

Coagulation factor XIII, a transglutaminase which stabilises blood clots by covalently cross-linking fibrin, is essential for normal haemostasis. FXIII deficiency results in a life-long bleeding disorder with added complications in wound healing and tissue repair. Sequence changes in the human FXIIIA gene, largely missense mutations, are primarily responsible for inherited FXIII deficiency. We have carried out molecular analysis of the FXIIIA gene in two unrelated FXIII deficient individuals and identified three splice site mutations; a g→a at the exon 6 acceptor splice site, a g→a at the exon 7 donor splice site and a coding sequence T→G at the exon 8 donor splice site. We have also examined the FXIIIA mRNA in these patients and find that each mutation gives rise to multiple transcripts which vary in their relative abundance. The precise molecular mechanisms which result in these variant transcripts, and their relative abundance in our FXIII deficient patients, are discussed.

• References

• 1 Bohn H. Comparative studies on the fibrin-stabilising factors from human plasma, platelets and placenta. Annals of the New York Acad of Sciences 1972; 202: 256-72.
• 2 Schwartz ML, Pizzo SV, Hill RL, McKee PA. Human factor XIII from plasma and platelets. J Biol Chem 1973; 248: 1395-407.
• 3 Takagi T, Doolittle RF. Amino acid sequence studies on factor XIII and the peptide released during its activation by thrombin. Biochemistry 1974; 13: 750-6.
• 4 Lorand L, Losowsky MS, Miloszewski K. Human factor XIII fibrin stabil-ising factor. Progress in Hemostasis and Thrombosis 1980; 5: 245-90.
• 5 Hansen MS. Fibronectin and coagulation factor XIII increase blood platelet adhesion to fibrin. Thrombosis Research 1984; 34: 551-6.
• 6 Bale MD, Mosher DF. Thrombospondin is a substrate for blood-coagulation factor-XIIIa. Biochemistry 1986; 25: 5667-73.
• 7 Miloszewski KJA, Losowsky MS. Fibrin stabilisation and factor XIII deficiency. In: Fibrinogen, Fibrin Stabilisation and Fibrinolysis. Francis L. ed. Ellis Horwood; 1988. pp 175-202.
• 8 Duckert F, Jung E, Shmerling DH. A hitherto undescribed congenital haemorrhagic diathesis probably due to fibrin stabilising factor deficiency. Thromb Diathesis Haemorrhage 1960; 5: 179-86.
• 9 Board PG, Losowsky MS, Miloszewski KJA. Factor XIII: Inherited and acquired deficiency. Blood 1993; 7: 229-42.
• 10 Aslam S, Poon MC, Yee VC, Bowen DJ, Standen GR. Factor XIIIA Calgary: a candidate missense mutation (Leu667Pro) in the beta barrel 2 domain of the factor XIIIA subunit. Br J Haematol 1995; 91: 452-7.
• 11 Anwar R, Stewart AD, Miloszewski KJA, Losowsky MS, Markham AF. Molecular basis of inherited factor XIII deficiency: identification of multiple mutations provides insights into protein function. Br J Haematol 1995; 91: 728-35.
• 12 Mikkola H, Yee VC, Syrjala M, Seitz R, Egbring R, Petrini P, Ljung R, Ingerslev J, Teller DC, Peltonen L, Palotie A. 4 novel mutations in deficiency of coagulation factor XIII – consequences to expression and structure of the A subunit. Blood 1996; 87: 141-51.
• 13 Kamura T, Okamura T, Murakawa M, Tsuda H, Teshima T, Shibuya T, Harada M, Niho Y. Deficiency of coagulation factor XIIIa subunit caused by the dinucleotide deletion at the 5’-end of exon 3. J Clin Invest 1992; 90: 315-9.
• 14 Aslam S, Bowen DJ, Mandalaki T, Gialeraki R, Standen GR. Factor XIII(A) subunit deficiency due to a homozygous 13-base pair deletion in exon 3 of the A subunit gene. Am J Haematol 1996; 53: 77-80.
• 15 Kangsadalampai S, Fargesberth A, Caglayan SH, Board PG. New mutations causing the premature termination of translation in the A subunit gene of coagulation factor XIII. Thromb Haemost 1996; 76: 139-42.
• 16 Inbal A, Yee VC, Kornbrot N, Zivelin A, Brenner B, Seligsohn U. Factor XIII deficiency due to a Leu660Pro mutation in the factor XIII subunit-a gene in three unrelated Palestinian Arab families. Thromb Haemost 1997; 77: 1062-7.
• 17 Aslam S, Yee VC, Narayanan S, Duraisamy G, Standen GR. Structural analysis of a missense mutation (Val414Phe) in the catalytic core domain of the factor XIIIA subunit. Br J Haematol 1997; 98: 346-52.
• 18 Anwar R, Miloszewski KJA, Markham AF. Identification of a large deletion, spanning exons 4 to 11 of the human FXIIIA gene, in a Factor XIII deficient family. Blood 1998; 91: 149-53.
• 19 Chomczynski P, Sacchi N. Single step method of RNA extraction by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-9.
• 20 Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A laboratory Manual: 2nd edit. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, New York: 1989
• 21 Vreken P, Niessen RWLM, Peters M, Schaap MCL, Zuithoff-Rijntjes JGM, Sturk A. A point mutation in an invariant splice acceptor site results in a decreased mRNA level in a patient with severe coagulation factor XIII subunit A deficiency. Thromb Haemost 1995; 74: 584-9.
• 22 Yee VC, Pedersen LC, Trong IL, Bishop PD, Stenkamp RE, Teller DC. Three-dimensional structure of a transglutaminase: human blood coagulation factor XIII. Proc Natl Acad Sci USA 1994; 91: 7296-300.
• 23 Pedersen LC, Yee VC, Bishop PD, Trong IL, Teller DC, Stenkamp RE. Transglutaminase factor XIII uses proteinase-like catalytic triad to cross-link macromolecules. Protein Sci 1994; 3: 1131-5.
• 24 Holliday R, Murray V. Specificity in splicing. BioEssays 1994; 16: 771-4.
• 25 Estes PA, Cooke NE, Liebhaber SA. A native RNA structure controls alternative splice site selection and generates two human growth hormone isoforms. J Biol Chem 1992; 267: 14902-8.
• 26 Eperon IC, Ireland DC, Smith RA, Mayeda A, Krainer AR. Pathways for selection of 5’ splice sites by U1 snRNPs and SF2/ASF. EMBO J 1993; 12: 3607-17.
• 27 Humphrey MB, Bryan J, Cooper TA, Berget SM. A 32-nucleotide exon-splicing enhancer regulates usage of competing 5’ splice sites in a differential internal exon. Mol Cell Biol 1995; 15: 3979-88.
• 28 Ichinose A, Davie EW. Characterisation of the gene for the a subunit of factor XIII (plasma transglutaminase), a blood coagulation factor. Proc Natl Acad Sci USA 1988; 85: 5829-33.
• 29 Mikkola H, Muszbek L, Laiho E, Syrjala M, Hamalainen E, Haramura G, Salmi T, Peltonen L, Palotie A. Molecular mechanism of a mild phenotype in coagulation factor XIII (FXIII) deficiency: A splicing mutation permitting partial correct splicing of FXIII a-subunit mRNA. Blood 1997; 89: 1279-87.
• 30 Suzuki K, Henke J, Iwata M, Henke L, Tsuji H, Fukunaga T, Ishimoto G, Szekelyi M, Ito S. Novel polymorphisms and haplotypes in the human coagulation factor XIII A-subunit gene. Hum Gen 1996; 98: 393-5.