title: Copy number variation of two separate regulatory regions upstream of SOX9 causes isolated 46,XY or 46,XX disorder of sex development.
abstract: BACKGROUND
  SOX9 mutations cause the skeletal malformation syndrome campomelic dysplasia in combination with XY sex reversal. Studies in mice indicate that SOX9 acts as a testis-inducing transcription factor downstream of SRY, triggering Sertoli cell and testis differentiation. An SRY-dependent testis-specific enhancer for Sox9 has been identified only in mice. A previous study has implicated copy number variations (CNVs) of a 78 kb region 517-595 kb upstream of SOX9 in the aetiology of both 46,XY and 46,XX disorders of sex development (DSD). We wanted to better define this region for both disorders.
  
  
  RESULTS
  By CNV analysis, we identified SOX9 upstream duplications in three cases of SRY-negative 46,XX DSD, which together with previously reported duplications define a 68 kb region, 516-584 kb upstream of SOX9, designated XXSR (XX sex reversal region). More importantly, we identified heterozygous deletions in four families with SRY-positive 46,XY DSD without skeletal phenotype, which define a 32.5 kb interval 607.1-639.6 kb upstream of SOX9, designated XY sex reversal region (XYSR). To localise the suspected testis-specific enhancer, XYSR subfragments were tested in cell transfection and transgenic experiments. While transgenic experiments remained inconclusive, a 1.9 kb SRY-responsive subfragment drove expression specifically in Sertoli-like cells.
  
  
  CONCLUSIONS
  Our results indicate that isolated 46,XY and 46,XX DSD can be assigned to two separate regulatory regions, XYSR and XXSR, far upstream of SOX9. The 1.9 kb SRY-responsive subfragment from the XYSR might constitute the core of the Sertoli-cell enhancer of human SOX9, representing the so far missing link in the genetic cascade of male sex determination.
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany Faculty of Biology, University of Freiburg, Freiburg, Germany.
authors.forename: Gwang-Jin
authors.initials: GJ
authors.lastname: Kim
authors.affiliation: Institute of Biochemistry, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Sock
authors.affiliation: Department of Cell and Molecular Biology, Institute of Biochemistry and Biotechnology, University of Braunschweig, Braunschweig, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Buchberger
authors.affiliation: Institute of Human Genetics, University of Ulm, Ulm, Germany.
authors.forename: Walter
authors.initials: W
authors.lastname: Just
authors.affiliation: Department of Pediatrics and Adolescent Medicine, University Hospital Ulm, Ulm, Germany.
authors.forename: Friederike
authors.initials: F
authors.lastname: Denzer
authors.affiliation: University Hospital for Children and Adolescents, University of Leipzig, Leipzig, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hoepffner
authors.affiliation: Department of Pediatrics, Cornell University, New York, USA.
authors.forename: James
authors.initials: J
authors.lastname: German
authors.affiliation: Department of Medical and Molecular Genetics, University of Birmingham School of Medicine, Institute of Biomedical Research, Birmingham, UK.
authors.forename: Trevor
authors.initials: T
authors.lastname: Cole
authors.affiliation: Department of Paediatric Oncology, Birmingham Children's Hospital, Birmingham, UK.
authors.forename: Jillian
authors.initials: J
authors.lastname: Mann
authors.affiliation: Pediatrix Medical Group of Ohio, Columbus, Ohio, USA.
authors.forename: John H
authors.initials: JH
authors.lastname: Seguin
authors.affiliation: Central Ohio Pediatric Endocrinology and Diabetes Services (COPED), Columbus, Ohio, USA.
authors.forename: William
authors.initials: W
authors.lastname: Zipf
authors.affiliation: National Centre for Medical Genetics, Our Lady's Hospital for Sick Children, Dublin, Ireland.
authors.forename: Colm
authors.initials: C
authors.lastname: Costigan
authors.affiliation: Klinik für Gynäkologie, Charité, Frauenklinik Campus Virchow-Klinikum, Berlin, Germany.
authors.forename: Hardi
authors.initials: H
authors.lastname: Schmiady
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Rostásy
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Mildred
authors.initials: M
authors.lastname: Kramer
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Kaltenbach
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Rösler
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany Area of Human DNA Variability, Centro de Genómica e Investigación Oncológica (GENYO), Pfizer-Universidad de Granada-Junta de Andalucia, Granada, Spain.
authors.forename: Ina
authors.initials: I
authors.lastname: Georg
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Elke
authors.initials: E
authors.lastname: Troppmann
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Anne-Christin
authors.initials: AC
authors.lastname: Teichmann
authors.affiliation: Department of Pediatrics, Centre for Pediatrics and Adolescent Medicine, University of Freiburg, Freiburg, Germany.
authors.forename: Anika
authors.initials: A
authors.lastname: Salfelder
authors.affiliation: Institute of Biochemistry, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Sebastian A
authors.initials: SA
authors.lastname: Widholz
authors.affiliation: Institute of Human Genetics, Westfälische Wilhelms Universität Münster, Münster, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Wieacker
authors.affiliation: Department of Pediatric and Adolescent Medicine, University of Lübeck, Lübeck, Germany.
authors.forename: Olaf
authors.initials: O
authors.lastname: Hiort
authors.affiliation: Dipartamento di Patologia Umana ed Ereditaria, Biologia Generale e Genetica Medica, Pavia, Italy.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Camerino
authors.affiliation: Dipartamento di Patologia Umana ed Ereditaria, Biologia Generale e Genetica Medica, Pavia, Italy.
authors.forename: Orietta
authors.initials: O
authors.lastname: Radi
authors.affiliation: Institute of Biochemistry, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Wegner
authors.affiliation: Department of Cell and Molecular Biology, Institute of Biochemistry and Biotechnology, University of Braunschweig, Braunschweig, Germany.
authors.forename: Hans-Henning
authors.initials: HH
authors.lastname: Arnold
authors.affiliation: Institute of Human Genetics, University of Freiburg, Freiburg, Germany.
authors.forename: Gerd
authors.initials: G
authors.lastname: Scherer
mesh_terms: D000818:Animals; D002460:Cell Line; D015331:Cohort Studies; D056915:DNA Copy Number Variations; D012734:Disorders of Sex Development / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D010375:Pedigree; D012045:Regulatory Sequences, Nucleic Acid; D055758:SOX9 Transcription Factor / Q000235:genetics*
chemical_list: D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human
keywords: SOX9; SRY; disorder of sex development; enhancer; long-range control