title: Screening of EDA1 gene in X-linked anhidrotic ectodermal dysplasia using DHPLC: identification of 14 novel mutations in Italian patients.
abstract: Mutations within EDA1 gene, which encodes for the ectodysplasin, cause X-linked anhidrotic ectodermal dysplasia. In this study, 23 Italian patients with anhidrotic ectodermal dysplasia were analyzed for mutations in EDA1 gene. We set up a rapid protocol through denaturing high-performance liquid chromatography, followed by sequencing, that allowed the characterization of 18 mutations, 14 novel and 4 recurrent: 8 missense mutations (p.L51Q, p.H54R, p.R156H twice, p.C332F, p.D316H, p.T378M, and p.A349T), 3 in-frame deletions (p.G82_P84del, p.A179_P191del, and p.L354del), 1 gross deletion (p.G168_G265del, identified through direct sequencing and PCR), 4 altered splicing (c.949-13T > C, c.741 + 1G/T, c.793 + 4A > T, and c.924 + 1G/T), 1 nonsense (p.Y3X), and 1 synonymous mutation (c.741G > A). Moreover, structural analysis of three missense mutations shows that alteration of the electrostatic surface of the protein (p.D316N), the break of intermonomer interactions (p.A349T) and destabilization of the single monomer structure (p.T378M), may irreversibly invalidate the EDA-A1 binding properties. Our data confirm and extend the large spectrum of EDA1 mutations and provide a rapid and efficient molecular protocol for testing EDA1 mutations in EDA patients.
authors.affiliation: Azienda Ospedaliera Universitaria Policlinico di Tor Vergata, Rome, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Conte
authors.forename: Stefano
authors.initials: S
authors.lastname: Gambardella
authors.forename: Cristina
authors.initials: C
authors.lastname: Bulli
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Rinaldi
authors.forename: Daniele
authors.initials: D
authors.lastname: Di Marino
authors.forename: Mattia
authors.initials: M
authors.lastname: Falconi
authors.forename: Placido
authors.initials: P
authors.lastname: Bramanti
authors.forename: Alessandro
authors.initials: A
authors.lastname: Desideri
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Novelli
mesh_terms: D017398:Alternative Splicing; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018389:Codon, Nonsense; D053358:Ectodermal Dysplasia 1, Anhidrotic / Q000235:genetics*; D053331:Ectodysplasins / Q000235:genetics*; D017353:Gene Deletion; D005820:Genetic Testing; D006801:Humans; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D007558:Italy; D008958:Models, Molecular; D009154:Mutation; D020125:Mutation, Missense; D055672:Static Electricity
chemical_list: D018389:Codon, Nonsense; D053331:Ectodysplasins