title: Molecular mechanism of a mild phenotype in coagulation factor XIII (FXIII) deficiency: a splicing mutation permitting partial correct splicing of FXIII A-subunit mRNA.
abstract: Congenital factor XIII (FXIII) deficiency is potentially a severe bleeding disorder, but in some cases, the symptoms may be fairly mild. In this study, we have characterized the molecular mechanism of a mild phenotype of FXIII A-subunit deficiency in a Finnish family with two affected sisters, one of whom has even had two successful pregnancies without regular substitution therapy. In the screening tests for FXIII deficiency, no A-subunit could be detected, but by using more sensitive assays, a minute amount of functional A-subunit was seen. 3H-putrescine incorporation assay showed distinct FXIII activity at the level of 0.35% of controls, and also the fibrin cross-linking pattern in the patients clotted plasma showed partial gamma-gamma dimerization. In Western blot analysis, a faint band of full-length FXIII A-subunit was detected in the patients' platelets. The patients have previously been identified as heterozygotes for the Arg661 --> Stop mutation. Here we report a T --> C transition at position +6 of intron C in their other allele. The transition affected splicing of FXIII mRNA resulting in low steady state levels of several variant mRNA transcripts. One transcript contained sequences of intron C, whereas two transcripts resulted from skipping of one or two exons. Additionally, correctly spliced mRNA lacking the Arg661 --> Stop mutation of the maternal allele could be detected. These results demonstrate that a mutation in splice donor site of intron C can result in several variant mRNA transcripts and even permit partial correct splicing of FXIII mRNA. Further, even the minute amount of correctly processed mRNA is sufficient for producing protein capable of gamma-gamma dimerization of fibrin. This is a rare example of an inherited functional human disorder in which a mutation affecting splicing still permits some correct splicing to occur and this has a beneficial effect to the phenotype of the patients.
authors.affiliation: Department of Clinical Chemistry, University of Helsinki, Finland.
authors.forename: H
authors.initials: H
authors.lastname: Mikkola
authors.forename: L
authors.initials: L
authors.lastname: Muszbek
authors.forename: E
authors.initials: E
authors.lastname: Laiho
authors.forename: M
authors.initials: M
authors.lastname: Syrjälä
authors.forename: E
authors.initials: E
authors.lastname: Hämäläinen
authors.forename: G
authors.initials: G
authors.lastname: Haramura
authors.forename: T
authors.initials: T
authors.lastname: Salmi
authors.forename: L
authors.initials: L
authors.lastname: Peltonen
authors.forename: A
authors.initials: A
authors.lastname: Palotie
mesh_terms: D004252:DNA Mutational Analysis; D019281:Dimerization; D005176:Factor XIII / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D005177:Factor XIII Deficiency / Q000097:blood / Q000235:genetics*; D005260:Female; D006579:Heterozygote; D006801:Humans; D007518:Isoantibodies / Q000276:immunology; D010375:Pedigree; D017354:Point Mutation; D018807:Polymorphism, Single-Stranded Conformational; D011247:Pregnancy; D011248:Pregnancy Complications; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*
chemical_list: D007518:Isoantibodies; D012333:RNA, Messenger; D005176:Factor XIII