Abstract
A gross deletion in the factor VIII (FVIII) mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR) for a patient with moderately severe hemophilia A. Sequencing of the RT-PCR product depicted a 177-bp deletion ranging from nucleotide (nt) 6724 to nt 6900 of FVIII cDNA, exactly corresponding to the whole exon 25. Further study of the genomic DNA revealed the presence of a single base pair substitution (G >A) at position –1 of intron 24. The absolute consensus AG doublet of the intron 24 splicing acceptor changed to AA. In the novel splice site mutation, exon 24 was erroneously spliced to exon 26, skipping exon 25. The FVIII antigen level was normal despite the markedly reduced functional activity. Since exon 25 corresponds to part of the C2 domain, we speculate that for this patient the aberrant C2 domain markedly reduces binding affinity of FVIII protein to the phospholipid membrane, thus severely impairing the protein function.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Kemball-Cook G, Tuddenham EGD, Wacey AI (1998) The factor VIII structure and mutation resource site: HAMSTeRs version 4. Nucleic Acids Res 26:216–219, available at http://europium.csc.mrc.ac.uk, accessed 20 October 2002
Waseem N, Bagnall R, Green PM, Gianelli F, the Haemophilia Centers (1999) Start of UK confidential haemophilia A database: analysis of 142 patients by solid phase fluorescent chemical cleavage of mismatch. Thromb Haemost 81:900–905
Naylor JA, Green PM, Montandon AJ, Rizza CR, Giannelli F (1991) Detection of three novel mutations in two hemophilia A patients by rapid screening of whole essential region of factor VIII gene. Lancet 337:635–639
Naylor JA, Green PM, Rizza CR, Giannelli F (1992) Factor VIII gene explains all cases of hemophilia A. Lancet 340:1066–1067
David D, Tavares A, Lavinha J (1995) Characterization of a splicing mutation in the factor VIII gene at the RNA level. Hum Genet 95:109–111
Youssoufian H, Kazazian HH Jr, Patel A, Aronis S, Tsiftis G, Hoyer LW, Antonarakis SE (1988) Mild hemophilia A associated with cryptic donor splice site mutation in intron 4 of the factor VIII gene. Genomics 2:32–36
Liu M-L, Murphy MEP, Thompson AR (1998) A domain mutations in 65 haemophilia A families and molecular modelling of dysfunctional factor VIII proteins. Br J Haematol 103:1051–1060
Pratt KP, Shen BW, Takeshima K, Davie EW, Fujikawa K, Stoddard BL (1999) Structure of the C2 domain of human factor VIII at 1.5A resolution. Nature 402:439–441
Liu M-L, Shen BW, Nakaya S, Pratt KP, Fujikawa K, Davie EW, Stoddard BL, Thompson AR (2000) Hemophilic factor VIII C1- and C2-domain missense mutations and their modeling to the 1.5-angstrom human C2-domain crystal structure. Blood 96:979–987
Pipe SW, Kaufman RJ (1996) Factor VIII C2 domain missense mutations exhibit defective trafficking of biologically functional proteins. J Biol Chem 271:25671–25676
Author information
Authors and Affiliations
Corresponding author
Additional information
An erratum to this article is available at http://dx.doi.org/10.1007/s00277-003-0657-6.
Rights and permissions
About this article
Cite this article
Gau, J.P., Hsu, H.C., Chau, W.K. et al. A novel splicing acceptor mutation of the factor VIII gene producing skipping of exon 25. Ann Hematol 82, 175–177 (2003). https://doi.org/10.1007/s00277-002-0592-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00277-002-0592-y