title: Spectrum of mutations in CRM-positive and CRM-reduced hemophilia A.
abstract: Hemophilia A is due to the functional deficiency of factor VII (FVIII, gene locus F8C). Although half the patients have no detectable FVIII protein in their plasma, the more rare patients (approximately 5%) have normal levels of a dysfunctional FVIII and are termed cross-reacting material (CRM)-positive. More commonly (approximately 45%), patients have plasma FVIII protein reduced to an extent roughly comparable to the level of FVIII activity and are designated CRM-reduced. We used denaturing gradient gel electrophoresis to screen for mutations within the F8C gene of 11 patients (6 CRM-positive, 5 CRM-reduced) and identified 9 different mutations in 9 patients after analyses of all 26 exons, the promoter region, and the polyadenylation site. Six mutations have not been described previously. Five were missense (Ser289Leu, Ser558Phe, Val634Ala, Val634-Met, Asn1441Lys), and the sixth was a 3-bp deletion (delta Phe652). A review of the literature and the assay of FVIII antigen in 5 hemophilia A patients with previously identified missense mutations from this laboratory yielded a total of 20 other unique CRM-reduced and CRM-positive mutations. Almost all CRM-positive/reduced mutations (24/26) were missense, and many (12/26) occurred at CpG dinucleotides. We examined 19 missense mutations for evolutionary conservation using the portions of the porcine and murine F8C sequences that are known, and 18/19 amino acid residues altered by mutation in these patients were conserved. Almost 50% of mutations (11/26) clustered in the A2 domain, suggesting that this region is critical for the function of FVIII.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Center for Medical Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: M J
authors.initials: MJ
authors.lastname: McGinniss
authors.forename: H H
authors.initials: HH
authors.lastname: Kazazian
authors.forename: L W
authors.initials: LW
authors.lastname: Hoyer
authors.forename: L
authors.initials: L
authors.lastname: Bi
authors.forename: H
authors.initials: H
authors.lastname: Inaba
authors.forename: S E
authors.initials: SE
authors.lastname: Antonarakis
mesh_terms: D000818:Animals; D000941:Antigens / Q000032:analysis; D001483:Base Sequence; D017124:Conserved Sequence; D003429:Cross Reactions; D004247:DNA; D005169:Factor VIII / Q000235:genetics* / Q000276:immunology; D006467:Hemophilia A / Q000235:genetics*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D013552:Swine
chemical_list: D000941:Antigens; D005169:Factor VIII; D004247:DNA