title: Disruption of SATB2 or its long-range cis-regulation by SOX9 causes a syndromic form of Pierre Robin sequence.
abstract: Heterozygous loss-of-function (LOF) mutations in the gene encoding the DNA-binding protein, SATB2, result in micrognathia and cleft palate in both humans and mice. In three unrelated individuals, we show that translocation breakpoints (BPs) up to 896 kb 3' of SATB2 polyadenylation site cause a phenotype which is indistinguishable from that caused by SATB2 LOF mutations. This syndrome comprises long nose, small mouth, micrognathia, cleft palate, arachnodactyly and intellectual disability. These BPs map to a gene desert between PLCL1 and SATB2. We identified three putative cis-regulatory elements (CRE1-3) using a comparative genomic approach each of which would be placed in trans relative to SATB2 by all three BPs. CRE1-3 each bind p300 and mono-methylated H3K4 consistent with enhancer function. In silico analysis suggested that CRE1-3 contain one or more conserved SOX9-binding sites, and this binding was confirmed using chromatin immunoprecipitation on cells derived from mouse embryonic pharyngeal arch. Interphase bacterial artificial chromosome fluorescence in situ hybridization measurements in embryonic craniofacial tissues showed that the orthologous region in mice exhibits Satb2 expression-dependent chromatin decondensation consistent with Satb2 being a target gene of CRE1-3. To assess their in vivo function, we made multiple stable reporter transgenic lines for each enhancer in zebrafish. CRE2 was shown to drive SATB2-like expression in the embryonic craniofacial region. This expression could be eliminated by mutating the SOX9-binding site of CRE2. These observations suggest that SATB2 and SOX9 may be acting together via complex cis-regulation to coordinate the growth of the developing jaw.
authors.affiliation: MRC Human Genetics Unit, MRC Institute of Genetic and Molecular Medicine, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Jacqueline K
authors.initials: JK
authors.lastname: Rainger
authors.forename: Shipra
authors.initials: S
authors.lastname: Bhatia
authors.forename: Hemant
authors.initials: H
authors.lastname: Bengani
authors.forename: Philippe
authors.initials: P
authors.lastname: Gautier
authors.forename: Joe
authors.initials: J
authors.lastname: Rainger
authors.forename: Matt
authors.initials: M
authors.lastname: Pearson
authors.forename: Morad
authors.initials: M
authors.lastname: Ansari
authors.forename: Jayne
authors.initials: J
authors.lastname: Crow
authors.forename: Felicity
authors.initials: F
authors.lastname: Mehendale
authors.forename: Bozena
authors.initials: B
authors.lastname: Palinkasova
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Dixon
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Thompson
authors.forename: Mar
authors.initials: M
authors.lastname: Matarin
authors.forename: Sanjay M
authors.initials: SM
authors.lastname: Sisodiya
authors.forename: Dirk A
authors.initials: DA
authors.lastname: Kleinjan
authors.forename: David R
authors.initials: DR
authors.lastname: Fitzpatrick
mesh_terms: D000328:Adult; D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D004843:Epistasis, Genetic; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D036961:Matrix Attachment Region Binding Proteins / Q000235:genetics*; D051379:Mice; D009154:Mutation; D010855:Pierre Robin Syndrome / Q000175:diagnosis* / Q000235:genetics; D050436:Regulatory Elements, Transcriptional; D055758:SOX9 Transcription Factor / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D055815:Young Adult; D015027:Zebrafish
chemical_list: D036961:Matrix Attachment Region Binding Proteins; C521297:SATB2 protein, human; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D014157:Transcription Factors