title: Hailey-Hailey disease: identification of novel mutations in ATP2C1 and effect of missense mutation A528P on protein expression levels.
abstract: ATP2C1, encoding the human secretory pathway Ca(2+)-ATPase (hSPCA1), was recently identified as the defective gene in Hailey-Hailey disease (HHD), an autosomal dominant skin disorder characterized by abnormal keratinocyte adhesion in the suprabasal layers of the epidermis. In this study, we used denaturing high-performance liquid chromatography to screen all 28 exons and flanking intron boundaries of ATP2C1 for mutations in 9 HHD patients. Nine different mutations were identified. Five of these mutations, including one nonsense, one deletion, two splice-site, and one missense mutation, have not been previously reported. Recently, functional analysis of a series of site-specific mutants, designed to mimic missense mutations found in ATP2C1, uncovered specific defects in Ca(2+) and/or Mn(2+) transport and protein expression in mutant hSPCA1 polypeptides. In order to investigate the molecular and physiological basis of HHD in the patient carrying missense mutation A528P, located in the putative nucleotide binding domain of the molecule, site-directed mutagenesis was employed to introduce this mutation into the wild-type ATP2C1 (hSPCA1) sequence. Functional analyses of HHD-mutant A528P demonstrated a low level of protein expression, despite normal levels of mRNA and correct targeting to the Golgi, suggesting instability or abnormal folding of the mutated hSPCA1 polypeptides. Analogous to conclusions drawn from our previous studies, these results further support the theory of haploinsufficiency as a prevalent mechanism for the dominant inheritance of HHD, by suggesting that the level of hSPCA1 in epidermal cells is critical.
authors.affiliation: The Wellcome Trust Centre For Human Genetics, University of Oxford, Oxford, UK.
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Fairclough
authors.forename: Lorne
authors.initials: L
authors.lastname: Lonie
authors.forename: Kurt
authors.initials: K
authors.lastname: Van Baelen
authors.forename: Marek
authors.initials: M
authors.lastname: Haftek
authors.forename: Colin S
authors.initials: CS
authors.lastname: Munro
authors.forename: Susan M
authors.initials: SM
authors.lastname: Burge
authors.forename: Alain
authors.initials: A
authors.lastname: Hovnanian
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000378:metabolism; D018389:Codon, Nonsense; D017353:Gene Deletion; D015870:Gene Expression; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D020125:Mutation, Missense; D016506:Pemphigus, Benign Familial / Q000235:genetics*; D022821:RNA Splice Sites / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D018389:Codon, Nonsense; D022821:RNA Splice Sites; D012333:RNA, Messenger; C403624:ATP2C1 protein, human; D000252:Calcium-Transporting ATPases