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 language | English |
|---|---|
| Pages (from-to) | 4760-4770 |
| Number of pages | 11 |
| Journal | EMBO Journal |
| Volume | 17 |
| Issue number | 16 |
| DOIs | |
| Publication status | Published - Aug 17 1998 |
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Keywords
- Coactivator
- Dominant negative
- Resistance to thyroid hormone
- Thyroid receptors
- Transactivation
ASJC Scopus subject areas
- Genetics
- Cell Biology
Cite this
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 journal › Article
}
TY - JOUR
T1 - 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
AU - Collingwood, Trevor N.
AU - Wagner, Richard
AU - Matthews, Clare H.
AU - Clifton-Bligh, Rory J.
AU - Gurnell, Mark
AU - Rajanayagam, Odelia
AU - Agostini, Maura
AU - Fletterick, Robert J.
AU - Beck-Peccoz, Paolo
AU - Reinhardt, Walter
AU - Binder, Gerhard
AU - Ranke, Michael B.
AU - Hermus, Ad
AU - Hesch, Rolf D.
AU - Lazarus, John
AU - Newrick, Paul
AU - Parfitt, Vernon
AU - Raggatt, Peter
AU - De Zegher, Francis
AU - Chatterjee, V. Krishna K
PY - 1998/8/17
Y1 - 1998/8/17
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.
AB - 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.
KW - Coactivator
KW - Dominant negative
KW - Resistance to thyroid hormone
KW - Thyroid receptors
KW - Transactivation
UR - http://www.scopus.com/inward/record.url?scp=0032541336&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032541336&partnerID=8YFLogxK
U2 - 10.1093/emboj/17.16.4760
DO - 10.1093/emboj/17.16.4760
M3 - Article
C2 - 9707435
AN - SCOPUS:0032541336
VL - 17
SP - 4760
EP - 4770
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 16
ER -