title: Functional characterization of GATA3 mutations causing the hypoparathyroidism-deafness-renal (HDR) dysplasia syndrome: insight into mechanisms of DNA binding by the GATA3 transcription factor.
abstract: The hypoparathyroidism-deafness-renal (HDR) dysplasia syndrome is an autosomal dominant disorder caused by mutations of the dual zinc finger transcription factor, GATA3. We investigated 21 HDR probands and 14 patients with isolated hypoparathyroidism for GATA3 abnormalities. Thirteen different heterozygous germline mutations were identified in patients with HDR. These consisted of three nonsense mutations, six frameshifting deletions, two frameshifting insertions, one missense (Leu348Arg) mutation and one acceptor splice site mutation. The splice site mutation was demonstrated to cause a pre-mRNA processing abnormality leading to the use of an alternative acceptor site 8 bp downstream of the normal site, resulting in a frameshift and prematurely terminated protein. Electrophoretic mobility shift assays (EMSAs) revealed three classes of GATA3 mutations: those that lead to a loss of DNA binding which represent over 90% of all mutations, and involved a loss of the carboxy-terminal zinc finger; those that resulted in a reduced DNA-binding affinity; and those (e.g. Leu348Arg) that did not alter DNA binding or the affinity but likely altered the conformational change that occurs during binding in the DNA major groove as predicted by a three-dimensional modeling. These results elucidate further the molecular mechanisms underlying the altered functions of mutants of this zinc finger transcription factor and their role in causing this developmental anomaly. No mutations were identified in patients with isolated hypoparathyroidism, thereby indicating that GATA3 abnormalities are more likely to result in two or more of the phenotypic features of the HDR syndrome and not in one, such as isolated hypoparathyroidism.
authors.affiliation: Academic Endocrine Unit, Nuffield Department of Clinical Medicine, Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Headington, Oxford OX3 7LJ, UK.
authors.forename: Asif
authors.initials: A
authors.lastname: Ali
authors.forename: Paul T
authors.initials: PT
authors.lastname: Christie
authors.forename: Irina V
authors.initials: IV
authors.lastname: Grigorieva
authors.forename: Brian
authors.initials: B
authors.lastname: Harding
authors.forename: Hilde
authors.initials: H
authors.lastname: Van Esch
authors.forename: S Faisal
authors.initials: SF
authors.lastname: Ahmed
authors.forename: Maria
authors.initials: M
authors.lastname: Bitner-Glindzicz
authors.forename: Eberhard
authors.initials: E
authors.lastname: Blind
authors.forename: Catherine
authors.initials: C
authors.lastname: Bloch
authors.forename: Patricia
authors.initials: P
authors.lastname: Christin
authors.forename: Peter
authors.initials: P
authors.lastname: Clayton
authors.forename: Jozef
authors.initials: J
authors.lastname: Gecz
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gilbert-Dussardier
authors.forename: Encarna
authors.initials: E
authors.lastname: Guillen-Navarro
authors.forename: Anna
authors.initials: A
authors.lastname: Hackett
authors.forename: Isil
authors.initials: I
authors.lastname: Halac
authors.forename: Geoffrey N
authors.initials: GN
authors.lastname: Hendy
authors.forename: Fiona
authors.initials: F
authors.lastname: Lalloo
authors.forename: Christoph J
authors.initials: CJ
authors.lastname: Mache
authors.forename: Zulf
authors.initials: Z
authors.lastname: Mughal
authors.forename: Albert C M
authors.initials: AC
authors.lastname: Ong
authors.forename: Choni
authors.initials: C
authors.lastname: Rinat
authors.forename: Nicholas
authors.initials: N
authors.lastname: Shaw
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Smithson
authors.forename: John
authors.initials: J
authors.lastname: Tolmie
authors.forename: Jacques
authors.initials: J
authors.lastname: Weill
authors.forename: M Andrew
authors.initials: MA
authors.lastname: Nesbit
authors.forename: Rajesh V
authors.initials: RV
authors.lastname: Thakker
mesh_terms: D000015:Abnormalities, Multiple / Q000235:genetics*; D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003638:Deafness / Q000235:genetics*; D005260:Female; D050990:GATA3 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007011:Hypoparathyroidism / Q000235:genetics*; D007223:Infant; D007231:Infant, Newborn; D007668:Kidney / Q000002:abnormalities*; D008297:Male; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D022821:RNA Splice Sites / Q000235:genetics; D013577:Syndrome
chemical_list: D004268:DNA-Binding Proteins; D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D022821:RNA Splice Sites