title: Deletion of long-range sequences at Sox10 compromises developmental expression in a mouse model of Waardenburg-Shah (WS4) syndrome.
abstract: The transcription factor SOX10 is mutated in the human neurocristopathy Waardenburg-Shah syndrome (WS4), which is characterized by enteric aganglionosis and pigmentation defects. SOX10 directly regulates genes expressed in neural crest lineages, including the enteric ganglia and melanocytes. Although some SOX10 target genes have been reported, the mechanisms by which SOX10 expression is regulated remain elusive. Here, we describe a transgene-insertion mutant mouse line (Hry) that displays partial enteric aganglionosis, a loss of melanocytes, and decreased Sox10 expression in homozygous embryos. Mutation analysis of Sox10 coding sequences was negative, suggesting that non-coding regulatory sequences are disrupted. To isolate the Hry molecular defect, Sox10 genomic sequences were collected from multiple species, comparative sequence analysis was performed and software was designed (ExactPlus) to identify identical sequences shared among species. Mutation analysis of conserved sequences revealed a 15.9 kb deletion located 47.3 kb upstream of Sox10 in Hry mice. ExactPlus revealed three clusters of highly conserved sequences within the deletion, one of which shows strong enhancer potential in cultured melanocytes. These studies: (i) present a novel hypomorphic Sox10 mutation that results in a WS4-like phenotype in mice; (ii) demonstrate that a 15.9 kb deletion underlies the observed phenotype and likely removes sequences essential for Sox10 expression; (iii) combine a novel in silico method for comparative sequence analysis with in vitro functional assays to identify candidate regulatory sequences deleted in this strain. These studies will direct further analyses of Sox10 regulation and provide candidate sequences for mutation detection in WS4 patients lacking a SOX10-coding mutation.
authors.affiliation: Geome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Antonellis
authors.forename: William R
authors.initials: WR
authors.lastname: Bennett
authors.forename: Trevelyan R
authors.initials: TR
authors.lastname: Menheniott
authors.forename: Arjun B
authors.initials: AB
authors.lastname: Prasad
authors.forename: Shih-Queen
authors.initials: SQ
authors.lastname: Lee-Lin
authors.forename: Eric D
authors.initials: ED
authors.lastname: Green
authors.forename: Derek
authors.initials: D
authors.lastname: Paisley
authors.forename: Robert N
authors.initials: RN
authors.lastname: Kelsh
authors.forename: William J
authors.initials: WJ
authors.lastname: Pavan
authors.forename: Andrew
authors.initials: A
authors.lastname: Ward
mesh_terms: D000465:Algorithms; D000818:Animals; D001483:Base Sequence / Q000235:genetics*; D015139:Blotting, Southern; D002478:Cells, Cultured; D017124:Conserved Sequence / Q000235:genetics; D004252:DNA Mutational Analysis; D004622:Embryo, Mammalian / Q000378:metabolism*; D040461:Gene Components; D018507:Gene Expression Regulation, Developmental; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D017403:In Situ Hybridization; D008156:Luciferases; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D055757:SOXE Transcription Factors; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion / Q000235:genetics*; D013045:Species Specificity; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes / Q000235:genetics; D014849:Waardenburg Syndrome / Q000235:genetics*
chemical_list: D006609:High Mobility Group Proteins; C527923:SOX10 protein, human; D055757:SOXE Transcription Factors; C527924:Sox10 protein, mouse; D014157:Transcription Factors; D008156:Luciferases