title: Mutation analysis of the entire keratin 5 and 14 genes in patients with epidermolysis bullosa simplex and identification of novel mutations.
abstract: Epidermolysis bullosa simplex is a group of blistering skin disorders caused by defects in one of the keratin genes, KRT5 and KRT14. Previously reported KRT5 and KRT14 mutations are clustered in several hotspots, namely the rod ends of the 1A and 2B domains and in the non-helical linker region L12. Therefore, genomic KRT5 and KRT14 mutation analysis was initially limited to these hotspots. In this study we describe the screening of nine EBS patients for mutations in the hotspots. In two patients, with the Koebner and the Weber-Cockayne subtypes of epidermolysis bullosa simplex respectively, we could, however, not identify any mutation in one of the hotspot domains of KRT5 and KRT14. Therefore, it appeared to be necessary to screen the entire genes for mutations. For KRT5, a complete genomic mutation detection system was previously described. We now developed a complete genomic mutation detection system for KRT14. For the amplification of the KRT14 genes, we make use of restriction sites to exempt the keratin 14 pseudogene sequence from polymerase chain reaction amplification. Using the complete genomic mutation detection system for both KRT5 and KRT14, we identified four novel KRT5 mutations (IVS1-1G>C, K404E, A438D, E475K), two of which are outside the KRT5 hotspot domains, and three novel KRT14 mutations (IVS4+1G>A, L408M, L130P).
authors.affiliation: Department of Medical Genetics, University of Groningen, Groningen, The Netherlands.
authors.forename: Petra H L
authors.initials: PH
authors.lastname: Schuilenga-Hut
authors.forename: Pieter v d
authors.initials: Pv
authors.lastname: Vlies
authors.forename: Marcel F
authors.initials: MF
authors.lastname: Jonkman
authors.forename: Esmé
authors.initials: E
authors.lastname: Waanders
authors.forename: Charles H C M
authors.initials: CH
authors.lastname: Buys
authors.forename: Hans
authors.initials: H
authors.lastname: Scheffer
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D002478:Cells, Cultured; D004252:DNA Mutational Analysis / Q000379:methods*; D016110:Epidermolysis Bullosa Simplex / Q000235:genetics*; D005260:Female; D006801:Humans; D053547:Keratin-14; D053555:Keratin-5; D015603:Keratinocytes / Q000473:pathology; D007633:Keratins / Q000235:genetics*; D008297:Male; D020125:Mutation, Missense / Q000235:genetics*; D010641:Phenotype
chemical_list: C507540:KRT14 protein, human; C503940:KRT5 protein, human; D053547:Keratin-14; D053555:Keratin-5; D007633:Keratins