title: Genomic architecture at the Incontinentia Pigmenti locus favours de novo pathological alleles through different mechanisms.
abstract: IKBKG/NEMO gene mutations cause an X-linked, dominant neuroectodermal disorder named Incontinentia Pigmenti (IP). Located at Xq28, IKBKG/NEMO has a unique genomic organization, as it is part of a segmental duplication or low copy repeat (LCR1-LCR2, >99% identical) containing the gene and its pseudogene copy (IKBKGP). In the opposite direction and outside LCR1, IKBKG/NEMO partially overlaps G6PD, whose mutations cause a common X-linked human enzymopathy. The two LCRs in the IKBKG/NEMO locus are able to recombine through non-allelic homologous recombination producing either a pathological recurrent exon 4-10 IKBKG/NEMO deletion (IKBKGdel) or benign small copy number variations. We here report that the local high frequency of micro/macro-homologies, tandem repeats and repeat/repetitive sequences make the IKBKG/NEMO locus susceptible to novel pathological IP alterations. Indeed, we describe the first two independent instances of inter-locus gene conversion, occurring between the two LCRs, that copies the IKBKGP pseudogene variants into the functional IKBKG/NEMO, causing the de novo occurrence of p.Glu390ArgfsX61 and the IKBKGdel mutations, respectively. Subsequently, by investigating a group of 20 molecularly unsolved IP subjects using a high-density quantitative polymerase chain reaction assay, we have identified seven unique de novo deletions varying from 4.8 to ∼115 kb in length. Each deletion removes partially or completely both IKBKG/NEMO and the overlapping G6PD, thereby uncovering the first deletions disrupting the G6PD gene which were found in patients with IP. Interestingly, the 4.8 kb deletion removes the conserved bidirectional promoterB, shared by the two overlapping IKBKG/NEMO and G6PD genes, leaving intact the alternative IKBKG/NEMO unidirectional promoterA. This promoter, although active in the keratinocytes of the basal dermal layer, is down-regulated during late differentiation. Genomic analysis at the breakpoint sites indicated that other mutational forces, such as non-homologous end joining, Alu-Alu-mediated recombination and replication-based events, might enhance the vulnerability of the IP locus to produce de novo pathological IP alleles.
authors: Institute of Genetics and Biophysics Adriano Buzzati-Traverso, IGB-CNR, Naples 80131, Italy.
authors: Francesca
authors: F
authors: Fusco
authors: Mariateresa
authors: M
authors: Paciolla
authors: Federico
authors: F
authors: Napolitano
authors: Alessandra
authors: A
authors: Pescatore
authors: Irene
authors: I
authors: D'Addario
authors: Elodie
authors: E
authors: Bal
authors: Maria Brigida
authors: MB
authors: Lioi
authors: Asma
authors: A
authors: Smahi
authors: Maria Giuseppina
authors: MG
authors: Miano
authors: Matilde Valeria
authors: MV
authors: Ursini
mesh_terms: D000483:Alleles; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D023281:Genomics; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D059765:Homologous Recombination; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics*; D007184:Incontinentia Pigmenti / Q000235:genetics* / Q000473:pathology*; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D008297:Male; D018895:Microsatellite Repeats; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011544:Pseudogenes / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics*
chemical_list: C491731:IKBKG protein, human; D012333:RNA, Messenger; D005954:Glucosephosphate Dehydrogenase; D051550:I-kappa B Kinase