title: Analysis of 18 novel mutations in the factor VIII gene.
abstract: We describe 18 novel mutations, unreported in the Haemophilia A mutation Databases, that have been identified in a cohort of unrelated, Italian patients affected with haemophilia A (HA). Screening of the factor VIII gene (FVIII) was performed using denaturing high-performance liquid chromatography (DHPLC) and direct sequencing. Eight mutations were characterized as non-missense alterations, and the remaining 10 were missense mutations. Heterozygosity for the identified mutations was observed in the female relatives of patients belonging to eight families with sporadic cases. In an attempt to understand better the causative effect of the mutations and the clinical variability of the patients, missense mutation consequences were investigated for: (1) the nature of the new amino acid; (2) the location of the substituted amino acid within crystallographic and theoretical models; and (3) the degree of conservation of the native residue in factor VIII (FVIII) protein and FVIII-related protein family aligned sequences. These research tools have provided evidence that the mutations we describe involve residues that were conserved, at least in FVIII proteins, in all the species we compared.
authors.affiliation: Haemostasis and Haemophilia Laboratory, IV Paediatric Department, G. Gaslini Institute, Genova, Italy. patriziabicocchi@ospedale-gaslini.ge.it
authors.forename: Maria P
authors.initials: MP
authors.lastname: Bicocchi
authors.forename: Mirella
authors.initials: M
authors.lastname: Pasino
authors.forename: Tiziana
authors.initials: T
authors.lastname: Lanza
authors.forename: Federico
authors.initials: F
authors.lastname: Bottini
authors.forename: Elio
authors.initials: E
authors.lastname: Boeri
authors.forename: Pier G
authors.initials: PG
authors.lastname: Mori
authors.forename: Angelo C
authors.initials: AC
authors.lastname: Molinari
authors.forename: Camillo
authors.initials: C
authors.lastname: Rosano
authors.forename: Maura
authors.initials: M
authors.lastname: Acquila
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis; D005169:Factor VIII / Q000235:genetics*; D005260:Female; D017353:Gene Deletion; D006467:Hemophilia A / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D008297:Male; D009154:Mutation; D020125:Mutation, Missense; D010641:Phenotype; D017354:Point Mutation
chemical_list: D005169:Factor VIII