A role for helix 3 of the TRβ ligand-binding domain in coactivator recruitment identified by characterization of a third cluster of mutations in resistance to thyroid hormone

Trevor N. Collingwood, Richard Wagner, Clare H. Matthews, Rory J. Clifton-Bligh, Mark Gurnell, Odelia Rajanayagam, Maura Agostini, Robert J. Fletterick, Paolo Beck-Peccoz, Walter Reinhardt, Gerhard Binder, Michael B. Ranke, Ad Hermus, Rolf D. Hesch, John Lazarus, Paul Newrick, Vernon Parfitt, Peter Raggatt, Francis De Zegher, V. Krishna K Chatterjee

Research output: Contribution to journalArticle

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Abstract

Resistance to thyroid hormone (RTH) has hitherto been associated with thyroid hormone β receptor (TRβ) mutations which cluster in two regions (αα 310-353 and αα 429-461) of the hormone-binding domain and closely approximate the ligand-binding cavity. Here, we describe a third cluster of RTH mutations extending from αα 234-282 which constitute a third boundary of the ligand pocket. One mutant, T277A, exhibits impaired transactivation which is disproportionate to its mildly reduced ligand affinity (K(a)). T3-dependent recruitment of coactivators (SRC-1, ACTR) by mutant receptor-RXR heterodimers was reduced in comparison with wild-type. Cotransfection of SRC-1 restored transactivation by T277A. In the TRβ crystal structure this helix 3 residue is surface-exposed and is in close proximity to residues L454 and E457 in helix 12 which are known to be critical for coactivator interaction, suggesting that they all constitute part of a receptor-coactivator interface. The transcriptional function of other mutants (A234T, R243W/Q, A268D, Δ276I, A279V, R282S) in this cluster correlated with their reduced K(a) and they inhibited wild-type TRβ action in a dominant negative manner. DNA binding, heterodimerization and corepressor recruitment were preserved in all mutants, signifying the importance of these attributes for dominant negative activity and correlating with the absence of natural mutations in regions bordering the third cluster which mediate these functions.

Original languageEnglish
Pages (from-to)4760-4770
Number of pages11
JournalEMBO Journal
Volume17
Issue number16
DOIs
Publication statusPublished - Aug 17 1998

Fingerprint

Thyroid Hormone Resistance Syndrome
Thyroid Hormone Receptors
Thyroid Hormones
Ligands
Mutation
Transcriptional Activation
Co-Repressor Proteins
Crystal structure
Hormones
DNA

Keywords

  • Coactivator
  • Dominant negative
  • Resistance to thyroid hormone
  • Thyroid receptors
  • Transactivation

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Collingwood, T. N., Wagner, R., Matthews, C. H., Clifton-Bligh, R. J., Gurnell, M., Rajanayagam, O., ... Chatterjee, V. K. K. (1998). A role for helix 3 of the TRβ ligand-binding domain in coactivator recruitment identified by characterization of a third cluster of mutations in resistance to thyroid hormone. EMBO Journal, 17(16), 4760-4770. https://doi.org/10.1093/emboj/17.16.4760

A role for helix 3 of the TRβ ligand-binding domain in coactivator recruitment identified by characterization of a third cluster of mutations in resistance to thyroid hormone. / Collingwood, Trevor N.; Wagner, Richard; Matthews, Clare H.; Clifton-Bligh, Rory J.; Gurnell, Mark; Rajanayagam, Odelia; Agostini, Maura; Fletterick, Robert J.; Beck-Peccoz, Paolo; Reinhardt, Walter; Binder, Gerhard; Ranke, Michael B.; Hermus, Ad; Hesch, Rolf D.; Lazarus, John; Newrick, Paul; Parfitt, Vernon; Raggatt, Peter; De Zegher, Francis; Chatterjee, V. Krishna K.

In: EMBO Journal, Vol. 17, No. 16, 17.08.1998, p. 4760-4770.

Research output: Contribution to journalArticle

Collingwood, TN, Wagner, R, Matthews, CH, Clifton-Bligh, RJ, Gurnell, M, Rajanayagam, O, Agostini, M, Fletterick, RJ, Beck-Peccoz, P, Reinhardt, W, Binder, G, Ranke, MB, Hermus, A, Hesch, RD, Lazarus, J, Newrick, P, Parfitt, V, Raggatt, P, De Zegher, F & Chatterjee, VKK 1998, 'A role for helix 3 of the TRβ ligand-binding domain in coactivator recruitment identified by characterization of a third cluster of mutations in resistance to thyroid hormone', EMBO Journal, vol. 17, no. 16, pp. 4760-4770. https://doi.org/10.1093/emboj/17.16.4760
Collingwood, Trevor N. ; Wagner, Richard ; Matthews, Clare H. ; Clifton-Bligh, Rory J. ; Gurnell, Mark ; Rajanayagam, Odelia ; Agostini, Maura ; Fletterick, Robert J. ; Beck-Peccoz, Paolo ; Reinhardt, Walter ; Binder, Gerhard ; Ranke, Michael B. ; Hermus, Ad ; Hesch, Rolf D. ; Lazarus, John ; Newrick, Paul ; Parfitt, Vernon ; Raggatt, Peter ; De Zegher, Francis ; Chatterjee, V. Krishna K. / A role for helix 3 of the TRβ ligand-binding domain in coactivator recruitment identified by characterization of a third cluster of mutations in resistance to thyroid hormone. In: EMBO Journal. 1998 ; Vol. 17, No. 16. pp. 4760-4770.
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AU - De Zegher, Francis

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N2 - Resistance to thyroid hormone (RTH) has hitherto been associated with thyroid hormone β receptor (TRβ) mutations which cluster in two regions (αα 310-353 and αα 429-461) of the hormone-binding domain and closely approximate the ligand-binding cavity. Here, we describe a third cluster of RTH mutations extending from αα 234-282 which constitute a third boundary of the ligand pocket. One mutant, T277A, exhibits impaired transactivation which is disproportionate to its mildly reduced ligand affinity (K(a)). T3-dependent recruitment of coactivators (SRC-1, ACTR) by mutant receptor-RXR heterodimers was reduced in comparison with wild-type. Cotransfection of SRC-1 restored transactivation by T277A. In the TRβ crystal structure this helix 3 residue is surface-exposed and is in close proximity to residues L454 and E457 in helix 12 which are known to be critical for coactivator interaction, suggesting that they all constitute part of a receptor-coactivator interface. The transcriptional function of other mutants (A234T, R243W/Q, A268D, Δ276I, A279V, R282S) in this cluster correlated with their reduced K(a) and they inhibited wild-type TRβ action in a dominant negative manner. DNA binding, heterodimerization and corepressor recruitment were preserved in all mutants, signifying the importance of these attributes for dominant negative activity and correlating with the absence of natural mutations in regions bordering the third cluster which mediate these functions.

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