Mechanisms of the Mouse Orphan Nuclear Receptor TR2-11-mediated Gene Suppression

The mouse orphan nuclear receptor TR2-11 functions as a repressor for reporter genes containing a direct repeat-5 or direct repeat-4 hormone response element. The functional domains responsible for its suppressive activity are defined, including the DNA-binding domain and the ligand-binding domain. The C-terminal 30 amino acid residues can be deleted without compromising its suppressive activity, whereas a deletion for 40 amino acids completely abolishes the suppressive activity and receptor dimerization, and reduces the DNA-binding affinity. Point mutation at three conserved leucine residues located on the predicted dimer interface abolishes the suppressive activity, receptor dimerization and its DNA binding property. However, mutation at two consecutive glutamate residues located within the hinge between the last two helices of the ligand-binding domain (helix 10 and helix 11 according to the human retinoid receptor X α structure) drastically reduces its DNA-binding affinity and abrogates the suppressive activity without compromising its ability to dimerize, indicating that receptor dimerization property can be functionally uncoupled from its suppressive activity. A transferable, active silencing activity is encoded within the DEF segment of the receptor molecule, as evidenced by the suppression of a GAL4 reporter by a chimeric protein containing the DNA-binding domain of GAL4 and the DEF segment of TR2-11. Moreover, the C-terminal 49 amino acid sequence is required for this trans-suppressive activity. It is suggested that TR2-11 functions as a repressor, mediated by mechanisms requiring high affinity DNA binding, receptor dimerization, and active silencing. The mouse orphan nuclear receptor TR2-11 functions as a repressor for reporter genes containing a direct repeat-5 or direct repeat-4 hormone response element. The functional domains responsible for its suppressive activity are defined, including the DNA-binding domain and the ligand-binding domain. The C-terminal 30 amino acid residues can be deleted without compromising its suppressive activity, whereas a deletion for 40 amino acids completely abolishes the suppressive activity and receptor dimerization, and reduces the DNA-binding affinity. Point mutation at three conserved leucine residues located on the predicted dimer interface abolishes the suppressive activity, receptor dimerization and its DNA binding property. However, mutation at two consecutive glutamate residues located within the hinge between the last two helices of the ligand-binding domain (helix 10 and helix 11 according to the human retinoid receptor X α structure) drastically reduces its DNA-binding affinity and abrogates the suppressive activity without compromising its ability to dimerize, indicating that receptor dimerization property can be functionally uncoupled from its suppressive activity. A transferable, active silencing activity is encoded within the DEF segment of the receptor molecule, as evidenced by the suppression of a GAL4 reporter by a chimeric protein containing the DNA-binding domain of GAL4 and the DEF segment of TR2-11. Moreover, the C-terminal 49 amino acid sequence is required for this trans-suppressive activity. It is suggested that TR2-11 functions as a repressor, mediated by mechanisms requiring high affinity DNA binding, receptor dimerization, and active silencing. Nuclear receptors constitute a large family of transcription factors that play key roles in gene regulation. Many of these receptors are transcription regulators that modulate target gene expression by binding to specific DNA sequences in its promoter region (1Wahli W. Martinez E. FASEB J. 1991; 5: 2243-2249Crossref PubMed Scopus (255) Google Scholar, 2Glass C.K. Endocrine Rev. 1994; 15: 391-407PubMed Google Scholar, 3Zhang X. Pfahl M. Trends Endocrinol. Metab. 1993; 4: 156-162Abstract Full Text PDF PubMed Scopus (110) Google Scholar, 4Evans R.M. Science. 1988; 240: 889-895Crossref PubMed Scopus (6341) Google Scholar, 5Beato M. Cell. 1989; 56: 335-344Abstract Full Text PDF PubMed Scopus (2852) Google Scholar, 6Mangelsdorf D.J. Thummel C. Beato M. Herrlich P. Schutz G. Umesono K. Blumberg B. Kastner P. Mark M. Chambon P. Evans R.M. Cell. 1995; 83: 835-839Abstract Full Text PDF PubMed Scopus (6110) Google Scholar). These include the receptors for glucocorticoid, estrogen, progesterone, vitamin D, thyroid hormone, and retinoic acid, as well as a large number of orphan members that have no known ligands. The nuclear receptors share a common modular structure consisting of a variable N terminus, a DNA-binding domain (DBD) 1The abbreviations used are: DBD, DNA-binding domain; DR, direct repeat; GFP, green fluorescent protein; GST, glutathione S-transferase; LBD, ligand-binding domain; N-CoR, nuclear receptor corepressor; PCR, polymerase chain reaction; RA, retinoic acid; RAR, retinoic acid receptor, RXR, retinoid receptor X; RARE, retinoic acid response element; SMRT, silencing mediator for retinoid and thyroid hormone receptor; TBP, TATA box-binding protein; tk; thymidine kinase. in the middle of the receptor molecule, and a C-terminal ligand-binding domain (LBD) (1Wahli W. Martinez E. FASEB J. 1991; 5: 2243-2249Crossref PubMed Scopus (255) Google Scholar, 2Glass C.K. Endocrine Rev. 1994; 15: 391-407PubMed Google Scholar). The LBD determines the ligand specificity by formation of a highly specified binding pocket and mediates receptor dimerization by interacting at the dimerization interface located on the LBD surface. In their apo-forms, nuclear receptors function as repressors by interacting with corepressors, such as the nuclear receptor corepressor (N-CoR) (7Horlein A.J. Naar A.M. Heinzel T. Torchia J. Gloss B. Kurokawa R. Ryan A. Kamei Y. Soderstrom M. Glass C.K. Rosenfeld M.G. Nature. 1995; 377: 397-403Crossref PubMed Scopus (1714) Google Scholar) and silencing mediator for retinoid and thyroid hormone receptors (SMRT) (8Chen J.D. Evans R.M. Nature. 1995; 377: 454-457Crossref PubMed Scopus (1715) Google Scholar). These corepressors interact with the apo-receptors at their hinge regions between the DBD and LBD. Ligand binding induces a conformational change of the receptor, resulting in the release of the corepressor and recruiting of the co-activator, which binds to the activation function-2 (AF-2) domain located at the C terminus of the receptor molecule (9Joyeux A. Cavailles V. Balaguer P. Nicolas J.C. Mol. Endocrinol. 1997; 11: 193-202Crossref PubMed Scopus (40) Google Scholar, 10Hong H. Kohli K. Garabedian M.J. Stallcup M. Mol. Cell Biol. 1997; 17: 2735-2744Crossref PubMed Scopus (497) Google Scholar, 11Voegel J.J. Heine M.J.S. Zechel C. Chambon P. Gronemeyer H. EMBO J. 1996; 15: 3667-3675Crossref PubMed Scopus (953) Google Scholar, 12Baur E.V. Zechel C. Heery D. Heine M.J.S. Garnier J.M. Vivat V. Douarin B.L. Gronemeyer H. Chambon P. Losson R. EMBO J. 1996; 15: 110-124Crossref PubMed Scopus (350) Google Scholar, 13Cavailles V. Dauvois S. L'Horset F. Lopez G. Hoare S. Kushner P. Parker M.G. EMBO J. 1995; 14: 3741-3751Crossref PubMed Scopus (673) Google Scholar, 14Thenot S. Henriquet C. Rochefort H. Cavailles V. J. Biol. Chem. 1997; 272: 12062-12068Abstract Full Text Full Text PDF PubMed Scopus (87) Google Scholar). Direct interaction with the components of the basal transcription machinery such as transcription factor IIB and TATA box-binding protein (TBP) has also been implicated in the ligand-independent silencing activity of some of these receptors (15Fondell J.D. Brunel F. Hisatake K. Roeder R.G. Mol. Cell. Biol. 1996; 16: 281-287Crossref PubMed Google Scholar, 16Baniahmad A. Ha I. Reinberg D. Tsai S. Tsai M.J. O'Malley B.W. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 8832-8836Crossref PubMed Scopus (301) Google Scholar). The mouse TR2-11 is a member of the orphan receptors and is expressed mainly in the developing germ cells in the testis (17Lee C.-H. Copeland N.G. Gilbert D.J. Jenkins N.A. Wei L.-N. Genomics. 1995; 30: 46-52Crossref PubMed Scopus (34) Google Scholar, 18Lee C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar). Several mRNA isoforms have been identified (19Lee C.-H. Chang L. Wei L.-N. J. Endocrinol. 1997; 152: 245-255Crossref PubMed Scopus (28) Google Scholar, 20Chang C. Kokontis J. Acakpo-Satchivi L. Liao S. Takeda H. Chang Y. Biochem. Biophys. Res. Commun. 1989; 165: 735-741Crossref PubMed Scopus (86) Google Scholar), including one that encodes the full-length receptor and an alternatively spliced isoform that encodes a truncated receptor deleted in the entire LBD (19Lee C.-H. Chang L. Wei L.-N. J. Endocrinol. 1997; 152: 245-255Crossref PubMed Scopus (28) Google Scholar). Our previous studies have shown that the full-length receptor strongly suppresses reporters containing either a direct repeat-5 C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar, C.-H. Chang L. Wei L.-N. J. Endocrinol. 1997; 152: 245-255Crossref PubMed Scopus (28) Google Scholar) from the human gene promoter Evans R.M. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar), or a response C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar) from the mouse retinoic gene promoter L. Wei L.-N. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). In the truncated receptor no in the of an the of the nuclear the full-length TR2-11 receptor is nuclear receptors to the large DEF segment of its region amino acid residues for as with amino acids for W. Vivat V. Chambon P. D. Gronemeyer H. Biol. 1996; PubMed Scopus Google Scholar). In to TR2-11 suppresses gene expression as by previous studies C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar, C.-H. Chang L. Wei L.-N. J. Endocrinol. 1997; 152: 245-255Crossref PubMed Scopus (28) Google Scholar, C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar, Chang C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar, Chang C. J. Biol. Chem. 1995; Full Text Full Text PDF Scopus Google Scholar), have to the functional domains responsible for its activity and to TR2-11 or mechanisms by receptors for a strongly suppressive activity. In this the mechanisms the suppressive activity of TR2-11 in the reporter C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar). In this has been shown that of the reporter by the full-length TR2-11 receptor, by the truncated the receptors by deletion and in their ability and DNA binding and these to their suppressive in the reporter the of the predicted within the LBD in receptor dimerization, DNA binding, and suppressive activity. identified one conserved glutamate located between helices 10 and which for the suppressive activity and DNA-binding for receptor on to its suppressive activity is by the GAL4 protein the of its interaction with the common corepressor as well as with The containing a as C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar). The reporter by of a promoter between and of the gene The H. M. P. H. A. Nature. PubMed Scopus Google Scholar) the in of a The containing the full-length TR2-11 amino acid for expression as C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). The amino acid according to previous C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar). the C-terminal of deleted in from the C terminus by polymerase chain by and and used to the of the expression and deleted for and 10 amino from the C deleted in the amino acid residues variable and deleted in the amino acids variable region and the Point a DNA Google Scholar). The and used to in the The at the consecutive three leucine residues by the sequence with a sequence The at the two glutamate residues at and by the sequence with a sequence fluorescent protein of and by the from the expression the in of the and the of the The DNA containing the of the human promoter with TR2-11 and its a The as C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). the in with for at in the of of and by in by the of receptor binding to the the of a and used to as C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). cells in with with the suppressive activity of TR2-11 and its the reporters or the receptor expression and an cells by C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). of expression of the and of the at the of in the and for of the and as C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). The specific reporter activity as by to the activity. in and three to to the and the of the the trans-suppressive activity of GAL4 and on a reporter containing of the at the of a thymidine as The expression of the and of reporter as GAL4 and the domain acids in to the entire DEF the DEF deleted in the C-terminal 49 amino and the amino acid containing a reporter by three of GAL4 binding the the GAL4 DNA-binding domain and the the activation domain of GAL4 from A of and by of the LBD of TR2-11 the and the The by or from the The and the full-length The and amino acids D. H. A. Evans R.M. Proc. Natl. Acad. Sci. U. S. A. 1995; PubMed Scopus Google Scholar). The amino acids (7Horlein A.J. Naar A.M. Heinzel T. Torchia J. Gloss B. Kurokawa R. Ryan A. Kamei Y. Soderstrom M. Glass C.K. Rosenfeld M.G. Nature. 1995; 377: 397-403Crossref PubMed Scopus (1714) Google Scholar) to the The TR2-11 include the full-length the DEF segment the DEF deleted in the C-terminal 49 amino acids and the two and The TR2-11 include the DEF segment a DEF segment acids deleted at its C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar), as well as the C-terminal of the DEF segment The and from The and of activity as C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). The full-length TR2-11 the for the of in a of of protein in or to a and TR2-11 protein in The binding and a for at C. The protein by with a of the binding without and The protein with glutathione in and by In previous have that TR2-11 a strongly suppressive activity on the reporters containing either a the human C.-H. Chang L. Wei L.-N. Mol. Reprod. Dev. 1996; 44: 305-314Crossref PubMed Google Scholar) or a hormone response the mouse C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar). In this have two reporter to the mechanisms of the suppressive activity of TR2-11. The a reporter that been used to mediated by a element. In to the of in this also a promoter from which this the a well gene to The H. M. P. H. A. Nature. PubMed Scopus Google Scholar). the functional domains of TR2-11 that required for its suppressive activity, by from the N and C of the receptor, and by specific in the LBD of TR2-11. A the of two C-terminal and used in these The suppressive activity of receptor on of the and the reporters in and the are shown in and with the with a expression expression of the TR2-11 or three C-terminal and in of on activity and on activity. In deletion of 49 amino acids from the C terminus completely the suppressive of TR2-11 on the acid deletion the receptor function in the reporter the receptor function in the reporter amino acids or also in the of the suppressive activity The deletion which the DBD suppressive whereas a deletion the DBD completely the suppressive activity these that the variable region and the C-terminal 30 amino acid sequence are required for the suppressive activity of TR2-11 on either the region between the and the amino acid from the C terminus between the two reporter whereas the DBD and the LBD are for the suppressive activity in reporter acid of TR2-11 and receptors a sequence in the region containing the which to helices according to the structure W. M. Chambon P. Gronemeyer H. D. Nature. 1995; PubMed Scopus Google Scholar). this a sequence located in the is highly The sequence as shown in and the receptor to the suppressive activity in reporter a function of these conserved leucine residues in the suppressive activity of TR2-11. The that deletion of 40 amino 30 amino from the C terminus of TR2-11 in a of the suppressive activity suggested an of the 10 amino acids the and the residues from the C in this receptor activity. nuclear receptors that this acid sequence the hinge region between helices 10 and as well as the of helix a glutamate in the hinge region conserved and the hinge region between helices 10 and 11 for the suppressive activity. The two consecutive glutamate residues to and as and in shown in and this mutation completely the suppressive activity in reporter which glutamate mutation as at and at that mutation at the suppressive activity. is that the conserved glutamate an in The of the receptors to an on these reporters a the of protein or in protein these receptors the cells and the by a as shown in D. The receptor with a nuclear which also for the receptors of C-terminal and In the receptor deleted in the N terminus and the DBD, which deleted in the nuclear a completely The deletion also a nuclear to the In protein of these as evidenced by the that the nuclear fluorescent be for the also the activity of the receptors with in the reporter and a of activity as that shown in and for these receptors these is that the which is deleted in the N terminus and the DBD, are and the is for the function of nuclear dimerization have been defined, one within the of the DBD and the within the LBD T. Umesono K. Evans R.M. Mol. Endocrinol. 1996; Google Scholar). to dimerization of TR2-11 in and dimerization also required for the suppressive activity of TR2-11. The ability of TR2-11 and its deletion to in the interaction of in the expression and by of the and the the shown in receptors containing the DEF segment of dimerization whereas the C-terminal 49 amino acids receptors to Moreover, the interaction between the receptors is the between and large a interaction between the dimer the sequence required for this from the C terminus the and in interaction as shown in B. In with the of their suppressive and receptors deleted for to 30 amino acids from the C terminus of interacting with the receptor, deletion to 40 amino acids drastically the interaction between the dimer is that deletion from the C-terminal 40 amino acids of TR2-11 abolishes receptor dimerization property and its ability to target gene In as shown in the and the completely their suppressive their ability to in the interaction shown in of the to interact with receptor of interacting with the receptor and the of interaction to that of the receptor these that dimerization is with the suppressive activity of TR2-11 evidenced by the of C-terminal deletion and the leucine mechanisms mediated by the conserved glutamate residues are also for the suppressive activity. TR2-11 in a DNA TR2-11 with the at the N terminus and to a The TR2-11 protein with in a and to the as and shown in TR2-11 from the the is that TR2-11 is of in a DNA the receptors that their dimerization property and suppressive activity, such as receptors deleted at the C terminus or at leucine or glutamate in the DNA binding the as the shown in the receptor to the to a with an at the by at and by a specific TR2-11 C. Chang L. X. C.-H. Wei L.-N. 1997; PubMed Scopus Google Scholar) to a with The completely its DNA binding property whereas the or the of binding at a affinity as by by a of the In to the of the the for the and by including in the as shown B. The specific protein of these indicating that the and also that the be by the DNA containing either one of the two and that containing one by TR2-11 no binding by TR2-11 It is that deletion or mutation from the C terminus of TR2-11 drastically its DNA-binding affinity. In to the of the by the mutation for DNA binding, to the binding affinity of this and the shown in a of the with of The of the and the an the of the from the The of the of the these as for the However, to the binding, the the receptor be is that TR2-11 binds as C-terminal deletion to 49 amino or the of binding to at a whereas mutation at the three leucine residues completely abolishes the DNA binding property. the suppressive activity of TR2-11 is and to the modular domain that is responsible for this activity, of its DEF segment to the in and and their activity on the reporter as and shown in the expression of in a suppression of the basal activity of the reporter as with the deletion of the C-terminal 49 amino acids this trans-suppressive activity whereas the C-terminal amino acids no trans-suppressive activity is that the suppressive activity of TR2-11 is and the LBD including the C-terminal segment last 49 amino It is known this C-terminal domain encodes a activation function the of for the transferable, suppressive of nuclear receptors receptor with the corepressors such as and (7Horlein A.J. Naar A.M. Heinzel T. Torchia J. Gloss B. Kurokawa R. Ryan A. Kamei Y. Soderstrom M. Glass C.K. Rosenfeld M.G. Nature. 1995; 377: 397-403Crossref PubMed Scopus (1714) Google Scholar, J.D. Evans R.M. Nature. 1995; 377: 454-457Crossref PubMed Scopus (1715) Google Scholar). with or has also been implicated in the function of nuclear receptors J.M. G. T. B. Pfahl M. Mol. Cell. Biol. 1993; PubMed Scopus Google Scholar). TR2-11 such a for its suppressive activity, the of TR2-11 interaction with the known corepressor or the interaction shown in the DEF region of TR2-11 to interact with the known receptor interacting domain of the the and the The for including N-CoR, and in and is that the suppressive activity of and requiring the LBD, is mediated by an interaction with the common receptor interacting domain of the or with or In this have the functional domains in the suppressive activity of the TR2-11 a deletion receptors by and their and including receptor dimerization and DNA these mutation is that receptor dimerization and DNA binding are for the suppressive activity of TR2-11 and the C-terminal 49 amino acid sequence is in the suppressive activity, receptor dimerization and DNA the of this region and The three leucine residues located on the predicted helix 10 are for receptor dimerization, DNA binding and suppressive activity. A glutamate located between helices 10 and which is conserved and is for the suppressive activity and DNA binding property for receptor activity. that the suppressive activity of TR2-11 is DNA can be functionally uncoupled from its activity. in the GAL4 reporter that TR2-11 encodes a suppressive domain in its LBD and the C-terminal 49 amino acid residues are required for this trans-suppressive activity. The of 40 amino acids between the and the reporter is It is that the conformational change of this receptor has a on the on the is also by that the C-terminal acid sequence an as evidenced by the of the this the of receptors between the two reporter the of the used in reporter is strongly by the the promoter from which this is to and has been as one of the retinoid receptors in such as and Evans R.M. Mol. 1995; PubMed Scopus Google Scholar). In the of expression is also shown to with A.M. 1997; PubMed Scopus Google Scholar). Our that suppression of on the gene to the expression of in developing germ The of TR2-11 to the is known to be that of the Chang C. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). has been suggested that for binding is the its suppressive activity on gene In this the of a trans-suppressive activity of in to its high affinity DNA It is suggested that suppression by TR2-11 at two by for DNA binding and by a transferable, active silencing activity. TR2-11 interact with or and the suppressive activity DNA binding, the can be The polymerase transcription TBP, has also been shown to interact with nuclear receptor (15Fondell J.D. Brunel F. Hisatake K. Roeder R.G. Mol. Cell. Biol. 1996; 16: 281-287Crossref PubMed Google Scholar, 16Baniahmad A. Ha I. Reinberg D. Tsai S. Tsai M.J. O'Malley B.W. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 8832-8836Crossref PubMed Scopus (301) Google Scholar). However, have to interaction of TR2-11 with this molecule in the interaction is that TR2-11 suppresses the target genes by with the transcription machinery such as However, with transcription factors be its DEF segment can be to domain and TR2-11 to be to function in a to that of the thyroid hormone receptor and However, interact with the known domain of the suppressive activity of TR2-11 is mediated by interaction with these domains of It to be of can interact with TR2-11. dimerization is for the suppressive activity evidenced by the of C-terminal and the dimerization is for this suppressive activity evidenced by of suppressive activity of the glutamate which of dimer also in receptors A. E. J. B. Mol. Cell. Biol. 1993; PubMed Scopus Google Scholar, G. Pfahl M. Mol. Cell. Biol. 1994; 14: PubMed Scopus Google Scholar), the which the conserved leucine residues is for dimerization and the suppressive activity. The C-terminal helix of TR2-11 is for its suppressive activity, the C-terminal amino acid region functions to the of the LBD and a specific to be for its high affinity DNA binding and suppressive activity, a mutation in this region completely abolishes the suppressive activity and drastically reduces its DNA-binding affinity. is also by the that the of domain and C terminus this domain to the active silencing. It be to the mutation the receptor The C-terminal acid sequence of the dimerization interface as well as the region of the which is for recruiting in receptor (9Joyeux A. Cavailles V. Balaguer P. Nicolas J.C. Mol. Endocrinol. 1997; 11: 193-202Crossref PubMed Scopus (40) Google Scholar, 10Hong H. Kohli K. Garabedian M.J. Stallcup M. Mol. Cell Biol. 1997; 17: 2735-2744Crossref PubMed Scopus (497) Google Scholar, 11Voegel J.J. Heine M.J.S. Zechel C. Chambon P. Gronemeyer H. EMBO J. 1996; 15: 3667-3675Crossref PubMed Scopus (953) Google Scholar, 12Baur E.V. Zechel C. Heery D. Heine M.J.S. Garnier J.M. Vivat V. Douarin B.L. Gronemeyer H. Chambon P. Losson R. EMBO J. 1996; 15: 110-124Crossref PubMed Scopus (350) Google Scholar, 13Cavailles V. Dauvois S. L'Horset F. Lopez G. Hoare S. Kushner P. Parker M.G. EMBO J. 1995; 14: 3741-3751Crossref PubMed Scopus (673) Google Scholar, 14Thenot S. Henriquet C. Rochefort H. Cavailles V. J. Biol. Chem. 1997; 272: 12062-12068Abstract Full Text Full Text PDF PubMed Scopus (87) Google Scholar). It this domain of TR2-11 an activation domain with a in In this in activation and the of as deletion of this domain in to and the receptor a to the to release the corepressor (8Chen J.D. Evans R.M. Nature. 1995; 377: 454-457Crossref PubMed Scopus (1715) Google Scholar, A. X. Tsai Tsai O'Malley Mol. Cell. Biol. 1995; 15: PubMed Google Scholar). It is suggested that a specific conformational change at the C terminus is by ligand binding, which the release of corepressors S. Mol. Cell. Biol. 1997; 17: PubMed Scopus Google Scholar). It is that TR2-11 also such a as a glutamate mutation is to release the suppressive activity. However, to be TR2-11 with corepressors and specific are for TR2-11 to function as an of a for in