title: Detection of missense mutations by single-strand conformational polymorphism (SSCP) analysis in five dysfunctional variants of coagulation factor VII.
abstract: Five unrelated subjects with dysfunctional coagulation factor VII (FVII) were studied in order to identify missense mutations affecting function. Exons 2 to 8 and the intron-exon junctions of their FVII genes were amplified from peripheral white blood cell DNA by PCR and screened by SSCP analysis. DNA fragments showing aberrant mobility were sequenced. The following mutations were identified: in case 1 (FVII:C < 1%, FVII:Ag 18%) a heterozygous A to G transition at nucleotide 8915 in exon 6 results in the amino acid substitution Lys-137 to Glu near the C-terminus of the FVIIa light chain; in case 2 (FVII:C 7%, FVII:Ag 47%) a heterozygous A to G transition at nucleotide 7834 in exon 5 results in the substitution of Gln-100 by Arg in the second EGF-like domain; in case 3 (FVII:C 20%, FVII:Ag 76%) a homozygous G to A transition at nucleotide position 6055 in exon 4 was detected resulting in substitution of Arg-79 by Gln in the first EGF-like domain; in case 5 (FVII:C 10%, FVII:Ag 52%) a heterozygous C to T transition at nucleotide position 6054 in exon 4 also results in the substitution of Arg79, but in this case it is replaced by Trp; case 4 (FVII:C < 1%, FVII:Ag 100%) was homozygous for a previously reported mutation (G to A) at nucleotide position 10715 in exon 8, substituting Gln for Arg at position 304 in the protease domain. Cases 1, 2 and 5 evidently have additional undetected mutations.
authors.affiliation: Haemostasis Research Group, Clinical Research Centre, Harrow, Middlesex, UK.
authors.forename: O
authors.initials: O
authors.lastname: Takamiya
authors.forename: G
authors.initials: G
authors.lastname: Kemball-Cook
authors.forename: D M
authors.initials: DM
authors.lastname: Martin
authors.forename: D N
authors.initials: DN
authors.lastname: Cooper
authors.forename: A
authors.initials: A
authors.lastname: von Felten
authors.forename: E
authors.initials: E
authors.lastname: Meili
authors.forename: I
authors.initials: I
authors.lastname: Hann
authors.forename: D R
authors.initials: DR
authors.lastname: Prangnell
authors.forename: H
authors.initials: H
authors.lastname: Lumley
authors.forename: E G
authors.initials: EG
authors.lastname: Tuddenham
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017124:Conserved Sequence; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis; D005167:Factor VII / Q000737:chemistry / Q000235:genetics*; D005168:Factor VII Deficiency / Q000235:genetics*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D005167:Factor VII; D004247:DNA