Pathogenic Effects of F8 Gene Mutations Identified in the Algerian Population Using In Silico Methods

Abstract

Since the publication of the F8 gene sequence, a large number of F8 gene mutations has been identified. These mutations are responsible for causing hemophilia A, an inherited bleeding disorder resulting from a reduced or absence of the coagulant FVIII protein activity. In the present study, we aimed to analyze the structural and functional impact of novel F8 gene mutations, previously identified in the Algerian population, through several in silico methods : HSF, ESE Finder, I-Mutant 2, SIFT, Polyphen 2, Align GVGD, ConSurf and Project Hope. Our analysis showed that the splice site mutation c.5219+1G>T is responsible of exon 14 skipping which cause the deletion of the protein B domain. Concerning the missense mutation c.200A> C (p.Lys48Thr), physicochemical changes between Lysine and Threonine were predicted to have deleterious effects on both structure and function of the protein. The c.200A> C introduces a neutral residue to the protein structure which could be responsible of hydrogen bonds loss causing improper folding of the FVIII protein. The novel missense mutation c.2189G> A (p.Cys711Tyr) is responsible of the rupture of the disulfide bridge (Cys630-Cys711) causing a FVIII protein destabilization. This study demonstrates the effectiveness of the combined use of several in silico methods to assess the deleterious effects of novel mutations. In silico methods, despite being useful in providing informations about effects of mutations, can also serve as a first-pass filter for experimental studies.