Human Molecular Genetics Advance Access originally published online on December 5, 2005
Human Molecular Genetics 2006 15(2):259-271; doi:10.1093/hmg/ddi442
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Deletion of long-range sequences at Sox10 compromises developmental expression in a mouse model of Waardenburg–Shah (WS4) syndrome
1Genome Technology Branch, 2NIH Intramural Sequencing Center (NISC) and 3Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA, 4Developmental Biology Programme and Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Bath, UK and 5Graduate Genetics Program, The George Washington University, Washington, DC 20052, USA
* To whom correspondence should be addressed at: Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, 49 Convent Dr., Building 49, Room 4A82, Bethesda, MD 20892, USA. Tel: +1 3014967584; Fax: +1 3014022170; Email: bpavan{at}nhgri.nih.gov
Received October 12, 2005; Accepted November 25, 2005
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.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
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  T. Werner, A. Hammer, M. Wahlbuhl, M. R. Bosl, and M. Wegner Multiple conserved regulatory elements with overlapping functions determine Sox10 expression in mouse embryogenesis Nucleic Acids Res., October 8, 2007; 35(19): 6526 - 6538. [Abstract] [Full Text] [PDF]
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