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Related Collections Hemostasis, Thrombosis, and Vascular Biology Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 1 October 2000, Vol. 96, No. 7, pp. 2496-2500 HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY Afibrinogenemia: first identification of a splicing mutation in the fibrinogen gamma chain gene leading to a major gamma chain truncation Rosanna Asselta, Stefano Duga, Tatjana Simonic, Massimo Malcovati, Elena Santagostino, Paul L. F. Giangrande, Pier Mannuccio Mannucci, and Maria Luisa Tenchini From the Department of Biology and Genetics for Medical Sciences, University of Milan; Institute of Veterinary Physiology and Biochemistry, University of Milan, Italy; Angelo Bianchi Bonomi Hemophilia and Thrombosis Center and Fondazione Luigi Villa, Department of Internal Medicine, University of Milan, Italy, and IRCCS Maggiore Hospital, Milan, Italy; and Oxford Haemophilia Centre, Churchill Hospital, Headington, Oxford, UK. Congenital afibrinogenemia is a rare autosomal recessive disorder characterized by the complete absence of plasma fibrinogen and by a bleeding tendency ranging from mild to moderately severe. Beside a deletion of the almost entire A-chain gene, only 2 missense mutations in the C-terminal domain of the B-chain have been very recently described as being associated with afibrinogenemia. We studied a Pakistani patient with unmeasurable plasma levels of functional and immunoreactive fibrinogen. Sequencing of the fibrinogen genes revealed a homozygous GA transition at position +5 of intron 1 of the -chain gene. The predicted mutant fibrinogen -chain would contain the signal peptide, followed by a short stretch of aberrant amino acids, preceding a premature stop codon. To demonstrate the causal role of the identified mutation, we prepared expression vectors containing a region of the fibrinogen -chain gene spanning from exon 1 to intron 4 and carrying either a G or an A at position +5 of intron 1. Transient transfection of the mutated plasmid in HeLa cells, followed by RNA extraction and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, allowed us to demonstrate the production of an erroneously spliced messenger RNA (mRNA), retaining intron 1, as shown by direct sequencing. A normal splicing occurred in HeLa cells transfected with the wild-type plasmid. This is the first report of a mutation in the fibrinogen -chain gene causing afibrinogenemia and indicates that, in addition to the A and B-chain genes, the -chain gene must also be considered in mutation screening for afibrinogenemia. © 2000 by The American Society of Hematology.   CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: C. Dall'Osso, I. Guella, S. Duga, N. Locatelli, E. M. Paraboschi, M. Spreafico, A. Afrasiabi, C. Pechlaner, F. Peyvandi, M. L. Tenchini, et al. Molecular characterization of three novel splicing mutations causing factor V deficiency and analysis of the F5 gene splicing pattern Haematologica, October 1, 2008; 93(10): 1505 - 1513. [Abstract] [Full Text] [PDF] S. Spena, M. L. Tenchini, and E. Buratti Cryptic splice site usage in exon 7 of the human fibrinogen B{beta}-chain gene is regulated by a naturally silent SF2/ASF binding site within this exon RNA, June 1, 2006; 12(6): 948 - 958. [Abstract] [Full Text] [PDF] P. M. Mannucci, S. Duga, and F. Peyvandi Recessively inherited coagulation disorders Blood, September 1, 2004; 104(5): 1243 - 1252. [Abstract] [Full Text] [PDF] C. Attanasio, A. David, and M. 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