X-ray structure of the orphan nuclear receptor RORbeta ligand-binding domain in the active conformation

Stehlin, Catherine; Wurtz, Jean‐Marie; Steinmetz, Anke; Greiner, Erich F.; Schüle, Roland; Moras, Dino; Renaud, Jean‐Paul

doi:10.1093/emboj/20.21.5822

Cited by 116 publications

(93 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Likewise, endogenous "fortuitously" bound LCFA does not stabilize the active conformation of the retinoic acid-related orphan receptor ligand binding domain (reviewed in Ref. 52). In contrast, the conformational responsiveness to LCFA-CoA of HNF-4␣ constructs containing the C-terminal F-domain was consistent with the paradigm regarding ligand-induced conformational change in ligand-responsive nuclear receptors, including the closely related RXR receptor as well as the retinoic acid receptor, progesterone receptor, estrogen receptor, vitamin D receptor, thyroid hormone receptor, androgen receptor, glucocorticoid receptor (reviewed in Ref.…”

Section: Discussionmentioning

confidence: 99%

Role of Regulatory F-domain in Hepatocyte Nuclear Factor-4α Ligand Specificity

Petrescu¹,

Hertz

Bar‐Tana

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Role of Regulatory F-domain in Hepatocyte Nuclear Factor-4α Ligand Specificity

Petrescu¹,

Hertz

Bar‐Tana

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

“…Several NRs previously referred to as orphan receptors have been shown to adopt the same regulatory paradigm, and physiologically relevant NR ligands now also include fatty acids (peroxisome proliferator-activated receptor), oxysterols (liver-X receptor), bile acids (farnesoid-X receptor), and xenobiotic compounds (pregnane-X receptor) (28,29). For some NRs, ligands are reported to serve an essential structural role, not a regulatory one (30)(31)(32)(33)(34)(35). The ligands for these receptors were first revealed by crystallographic studies highlighting a new structural genomics approach to the identification of ligands for orphan NRs (36).…”

mentioning

confidence: 99%

The crystal structures of human steroidogenic factor-1 and liver receptor homologue-1

Wang¹,

Zhang²,

Marimuthu³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

133

134

View full text Add to dashboard Cite

Steroidogenic factor-1 (SF-1) and liver receptor homologue-1 (LRH-1) belong to the fushi tarazu factor 1 subfamily of nuclear receptors. SF-1 is an essential factor for sex determination during development and regulates adrenal and gonadal steroidogenesis in the adult, whereas LRH-1 is a critical factor for development of endodermal tissues and regulates cholesterol and bile acid homeostasis. Regulatory ligands are unknown for SF-1 and LRH-1. A reported mouse LRH-1 structure revealed an empty pocket in a region commonly occupied by ligands in the structures of other nuclear receptors, and pocket-filling mutations did not alter the constitutive activity observed. Here we report the crystal structures of the putative ligand-binding domains of human SF-1 at 2.1-Å resolution and human LRH-1 at 2.5-Å resolution. Both structures bind a coactivator-derived peptide at the canonical activationfunction surface, thus adopting the transcriptionally activating conformation. In human LRH-1, coactivator peptide binding also occurs to a second site. We discovered in both structures a phospholipid molecule bound in a pocket of the putative ligand-binding domain. MS analysis of the protein samples used for crystallization indicated that the two proteins associate with a range of phospholipids. Mutations of the pocket-lining residues reduced the transcriptional activities of SF-1 and LRH-1 in mammalian cell transfection assays without affecting their expression levels. These results suggest that human SF-1 and LRH-1 may be ligand-binding receptors, although it remains to be seen if phospholipids or possibly other molecules regulate SF-1 or LRH-1 under physiological conditions.x-ray crystallography ͉ phospholipid ͉ nuclear receptor ͉ steroid ͉ bile S teroidogenic factor-1 (SF-1; AD4BP͞NR5A1) and liver receptor homologue-1 (LRH-1; CPF͞FTF͞NR5A2), expressed in man, are homologues of the fushi tarazu factor-1 of Drosophila (1) and FF1B of fish (2). Together, these factors constitute the NR5A subfamily of nuclear receptors (NRs) (3). SF-1 and LRH-1 function as monomers (4) to regulate genes by binding to similar response elements.SF-1 is expressed in the adrenal, testes, ovary, pituitary, hypothalamus, spleen, and skin and regulates genes that direct biosynthesis of adrenal and gonadal steroids as well as Mullerian hormone and gonadotropins (5, 6). SF-1 is essential for normal adrenal and gonadal development given that SF-1 knockout in mice causes adrenal and gonadal agenesis and impaired gonadotropin expression, resulting in postnatal death due to severe adrenal insufficiency (7,8). SF-1 knockout also causes abnormalities of the ventromedial hypothalamic nucleus, the control center for satiety and feeding, which suggests that SF-1 may have broader roles in the control of metabolism and obesity (9). In humans, partial loss-of-function mutations in SF-1 result in XY sex reversal and adrenal failure (10, 11). Although SF-1 is expressed in the ovary (12), a mutation of SF-1 was observed not to affect ovarian development; thus, SF-1 may no...

show abstract

“…ROR␣ is a potent transcriptional activator even in the absence of exogenously added ligand. However, the recent resolution of the crystal structures of ROR␣ and ROR␤ ligand-binding domains (LBDs), in combination with mutagenesis assays of the ROR␣ LBD, suggest that members of the ROR family require that their LBD be occupied by a ligand for transcriptional activation to occur (31)(32)(33). Regulation of ROR␣ transcriptional activity is also mediated through co-regulator recruitment.…”

mentioning

confidence: 99%

The Co-repressor Hairless Protects RORα Orphan Nuclear Receptor from Proteasome-mediated Degradation

Moraitis

Giguère

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

ROR␣ is a constitutively active orphan nuclear receptor essential for cerebellar development and is previously shown to regulate genes involved in both myogenesis and adipogenesis. The transcriptional activity of ROR␣ is dependent on the presence of a ubiquitous ligand and can be abolished by interaction with Hairless (Hr), a ligand-oblivious nuclear receptor co-repressor. In this study, we first demonstrate that ROR␣ is a shortlived protein and that treatment with the MG-132 proteasome inhibitor results in the accumulation of ubiquitin-conjugated receptor and inhibition of transcription. These data show that ROR␣ transcriptional activity and degradation are intrinsically linked. In addition, the introduction of inactivation mutations in the ligand-binding pocket and co-regulator-binding surface of ROR␣ significantly increases protein stability, indicating that ligand and/or co-regulator binding perpetuates ROR␣ degradation. Strikingly, expression of the co-repressor Hr results in the stabilization of ROR␣ because of an inhibition of proteasome-mediated degradation of the receptor. Stabilization of ROR␣ by Hr requires intact nuclear receptor recognition LXXLL motifs within Hr. Interestingly, the co-repressor nuclear receptor co-repressor (NCoR) has no effect on ROR␣ protein turnover. This study shows that stabilization of ROR␣ is an essential component of Hr-mediated repression and suggests a molecular mechanism to achieve transcriptional repression by a liganded receptor-co-repressor complex.The ubiquitin-proteasome pathway is the major system employed by eukaryotes for the selective degradation of cellular proteins that play key roles in cellular processes such as cell cycle regulation, differentiation, signal transduction, transcription, and chromosomal stabilization (reviewed in Refs. 1 and 2). Proteolytic degradation by the ubiquitin-proteasome system involves ATP-dependent covalent attachment of a macromolecular chain of ubiquitin (Ub) 1 molecules to the target protein, followed by degradation through the multicatalytic 26 S proteasome. The conjugation of Ub, a highly conserved 8.6-kDa protein, to its target protein is mediated by the serial action of three enzymes: E1, the Ub-activating enzyme, activates Ub in an ATP-dependent manner; E2, the Ub-conjugating enzyme, catalyzes the attachment of Ub to the substrate protein; and E3, the Ub-ligases, serves as a scaffold between E2 and the substrate and provides recognition specificity of the substrate. Ubiquitinylation of a substrate is reversible, and Ub moieties can be cleaved from a target protein by deubiquitinating enzymes. These enzymes assure that the cell is not depleted of a Ub pool. A protein tagged with a polyubiquitin chain is recognized and degraded by the 26 S proteasome complex. This complex is composed of a 19 S regulatory subcomplex, consisting of a "lid" subunit and a "base" subunit, the latter containing the six ATPases required for the degradation executed by the 20 S catalytic subcomplex (2).The Ub-proteasome pathway has recently emerged a...

show abstract

X-ray structure of the orphan nuclear receptor RORbeta ligand-binding domain in the active conformation

Cited by 116 publications

References 61 publications

Role of Regulatory F-domain in Hepatocyte Nuclear Factor-4α Ligand Specificity

Role of Regulatory F-domain in Hepatocyte Nuclear Factor-4α Ligand Specificity

The crystal structures of human steroidogenic factor-1 and liver receptor homologue-1

The Co-repressor Hairless Protects RORα Orphan Nuclear Receptor from Proteasome-mediated Degradation

Contact Info

Product

Resources

About