Characterization of GATA3 mutations in the hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome

M. Andrew Nesbit, Michael R. Bowl, Brian Harding, Asif Ali, Alejandro R Ayala, Carol Crowe, Angus Dobbie, Geeta Hampson, Ian Holdaway, Michael A. Levine, Robert McWilliams, Susan Rigden, Julian Sampson, Andrew J. Williams, Rajesh V. Thakker

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

The hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome is an autosomal dominant disorder caused by mutations of the dual zinc finger transcription factor, GATA3. The C-terminal zinc finger (ZnF2) binds DNA, whereas the N-terminal finger (ZnF1) stabilizes this DNA binding and interacts with other zinc finger proteins, such as the Friends of GATA (FOG). We have investigated seven HDR probands and their families for GATA3 abnormalities and have identified two nonsense mutations (Glu-228 → Stop and Arg-367 → Stop); two intragenic deletions that result in frameshifts from codons 201 and 355 with premature terminations at codons 205 and 370, respectively; one acceptor splice site mutation that leads to a frameshift from codon 351 and a premature termination at codon 367; and two missense mutations (Cys-318 → Arg and Asn-320 → Lys). The functional effects of these mutations, together with a previously reported GATA3 ZnF1 mutation and seven other engineered ZnF1 mutations, were assessed by electrophoretic mobility shift, dissociation, yeast two-hybrid and glutathione S-transferase pull-down assays. Mutations involving GATA3 ZnF2 or adjacent basic amino acids resulted in a loss of DNA binding, but those of ZnF1 either lead to a loss of interaction with specific FOG2 ZnFs or altered DNA-binding affinity. These findings are consistent with the proposed three-dimensional model of ZnF1, which has separate DNA and protein binding surfaces. Thus, our results, which expand the spectrum of HDR-associated GATA3 mutations and report the first acceptor splice site mutation, help to elucidate the molecular mechanisms that alter the function of this zinc finger transcription factor and its role in causing this developmental anomaly.

Original languageEnglish
Pages (from-to)22624-22634
Number of pages11
JournalJournal of Biological Chemistry
Volume279
Issue number21
DOIs
StatePublished - May 21 2004
Externally publishedYes

Fingerprint

Hypoparathyroidism
Deafness
Zinc
Kidney
RNA Splice Sites
Mutation
DNA
Zinc Fingers
GATA3 Transcription Factor
Nonsense Codon
Basic Amino Acids
Electrophoretic mobility
DNA-Binding Proteins
Glutathione Transferase
Codon
Yeast
Assays
Transcription Factors
Missense Mutation
Fingers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of GATA3 mutations in the hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome. / Nesbit, M. Andrew; Bowl, Michael R.; Harding, Brian; Ali, Asif; Ayala, Alejandro R; Crowe, Carol; Dobbie, Angus; Hampson, Geeta; Holdaway, Ian; Levine, Michael A.; McWilliams, Robert; Rigden, Susan; Sampson, Julian; Williams, Andrew J.; Thakker, Rajesh V.

In: Journal of Biological Chemistry, Vol. 279, No. 21, 21.05.2004, p. 22624-22634.

Research output: Contribution to journalArticle

Nesbit, MA, Bowl, MR, Harding, B, Ali, A, Ayala, AR, Crowe, C, Dobbie, A, Hampson, G, Holdaway, I, Levine, MA, McWilliams, R, Rigden, S, Sampson, J, Williams, AJ & Thakker, RV 2004, 'Characterization of GATA3 mutations in the hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome', Journal of Biological Chemistry, vol. 279, no. 21, pp. 22624-22634. https://doi.org/10.1074/jbc.M401797200
Nesbit, M. Andrew ; Bowl, Michael R. ; Harding, Brian ; Ali, Asif ; Ayala, Alejandro R ; Crowe, Carol ; Dobbie, Angus ; Hampson, Geeta ; Holdaway, Ian ; Levine, Michael A. ; McWilliams, Robert ; Rigden, Susan ; Sampson, Julian ; Williams, Andrew J. ; Thakker, Rajesh V. / Characterization of GATA3 mutations in the hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 21. pp. 22624-22634.
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AU - Harding, Brian

AU - Ali, Asif

AU - Ayala, Alejandro R

AU - Crowe, Carol

AU - Dobbie, Angus

AU - Hampson, Geeta

AU - Holdaway, Ian

AU - Levine, Michael A.

AU - McWilliams, Robert

AU - Rigden, Susan

AU - Sampson, Julian

AU - Williams, Andrew J.

AU - Thakker, Rajesh V.

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N2 - The hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome is an autosomal dominant disorder caused by mutations of the dual zinc finger transcription factor, GATA3. The C-terminal zinc finger (ZnF2) binds DNA, whereas the N-terminal finger (ZnF1) stabilizes this DNA binding and interacts with other zinc finger proteins, such as the Friends of GATA (FOG). We have investigated seven HDR probands and their families for GATA3 abnormalities and have identified two nonsense mutations (Glu-228 → Stop and Arg-367 → Stop); two intragenic deletions that result in frameshifts from codons 201 and 355 with premature terminations at codons 205 and 370, respectively; one acceptor splice site mutation that leads to a frameshift from codon 351 and a premature termination at codon 367; and two missense mutations (Cys-318 → Arg and Asn-320 → Lys). The functional effects of these mutations, together with a previously reported GATA3 ZnF1 mutation and seven other engineered ZnF1 mutations, were assessed by electrophoretic mobility shift, dissociation, yeast two-hybrid and glutathione S-transferase pull-down assays. Mutations involving GATA3 ZnF2 or adjacent basic amino acids resulted in a loss of DNA binding, but those of ZnF1 either lead to a loss of interaction with specific FOG2 ZnFs or altered DNA-binding affinity. These findings are consistent with the proposed three-dimensional model of ZnF1, which has separate DNA and protein binding surfaces. Thus, our results, which expand the spectrum of HDR-associated GATA3 mutations and report the first acceptor splice site mutation, help to elucidate the molecular mechanisms that alter the function of this zinc finger transcription factor and its role in causing this developmental anomaly.

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