title: Molecular analysis of the genetic defect in a large cohort of IP patients and identification of novel NEMO mutations interfering with NF-kappaB activation.
abstract: Incontinentia Pigmenti (IP) is an X-linked genodermatosis that is lethal for males and present in females with abnormal skin pigmentation and high variable clinical signs, including retinal detachment, anodontia, alopecia, nail dystrophy and nervous system defects. The NF-kappaB essential modulator (NEMO) gene, responsible for IP, encodes the regulatory subunit of the IkappaB kinase (IKK) complex required for nuclear factor kappaB (NF-kappaB) activation. We analyzed the NEMO gene in 122 IP patients and identified mutations in 83 (36 familiar and 47 sporadic cases). The recurrent NEMO exon 4-10 deletion that is the major cause of the disease was present in 73 females (59.8%). In addition 10 point alterations (8.2% of females) were identified: three frameshift, three nonsense, three missense and one in-frame deletion of a single amino acid. We measured the effects of these NEMO point-mutations on NF-kappaB signaling in nemo(-/-) deficient murine pre-B cells. A mutation in the N-terminal domain, required for IKK assembly, reduced but did not abolish NF-kappaB activation following lipopolysaccharide stimulation. Mutations that disrupt the C-terminal domain, required for the recruitment of upstream factors, showed lower or no NF-kappaB activation. A phenotype score based on clinical features of our IP patients was applied for summarizing disease severity. The score did not correlate with mutation type or domain affected indicating that other factors influence the severity of IP. Such a factor is likely to be X-inactivation. Indeed, 64% of our patients have extremely skewed X-inactivation pattern (>/=80 : 20). Overall IP pathogenesis thus depends on a combination of X-inactivation and protein domain that recruit upstream factors and activate NF-kappaB.
authors.affiliation: Institute of Genetics and Biophysics, Adriano Buzzati Traverso-CR, Naples, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Fusco
authors.forename: Tiziana
authors.initials: T
authors.lastname: Bardaro
authors.forename: Giorgia
authors.initials: G
authors.lastname: Fimiani
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Mercadante
authors.forename: Maria Giuseppina
authors.initials: MG
authors.lastname: Miano
authors.forename: Geppino
authors.initials: G
authors.lastname: Falco
authors.forename: Alain
authors.initials: A
authors.lastname: Israël
authors.forename: Gilles
authors.initials: G
authors.lastname: Courtois
authors.forename: Michele
authors.initials: M
authors.lastname: D'Urso
authors.forename: Matilde Valeria
authors.initials: MV
authors.lastname: Ursini
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D004303:Dosage Compensation, Genetic; D005091:Exons / Q000235:genetics; D040461:Gene Components; D005816:Genetic Complementation Test; D006801:Humans; D051550:I-kappa B Kinase; D047468:Immunoprecipitation; D007184:Incontinentia Pigmenti / Q000235:genetics*; D008070:Lipopolysaccharides; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics*; D016328:NF-kappa B / Q000378:metabolism*; D010375:Pedigree; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D014162:Transfection
chemical_list: D002352:Carrier Proteins; C491731:IKBKG protein, human; D008070:Lipopolysaccharides; D016328:NF-kappa B; D051550:I-kappa B Kinase