LXXLL-related Motifs in Dax-1 Have Target Specificity for the Orphan Nuclear Receptors Ad4BP/SF-1 and LRH-1

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2002 Dec 17

PMID 12482977

Citations 41

Authors

Taiga Suzuki

Megumi Kasahara

Hidefumi Yoshioka

Ken-Ichirou Morohashi

Kazuhiko Umesono

Affiliations

Soon will be listed here.

Abstract

The orphan receptor Ad4BP/SF-1 (NR5A1) is a constitutive activator, and its activity is repressed by another orphan receptor, Dax-1 (NR0B1). In the present study, we investigated the molecular mechanisms underlying this repression by Dax-1. Yeast two-hybrid and transient-transfection assays confirmed the necessity of three LXXLL-related motifs in Dax-1 for interaction with and repression of Ad4BP/SF-1. In vitro pull-down experiments confirmed that Dax-1 interacts with Ad4BP/SF-1 and also with LRH-1 (NR5A2). The target specificity of the LXXLL-related motifs was indicated by the observations that Ad4BP/SF-1, ERalpha (NR3A1), LRH-1, ERR2 (NR3B2), and fly FTZ-F1 (NR5A3) interacted through their ligand binding domains with all the LXXLL-related motifs in Dax-1 whereas HNF4 (NR2A1) and RORalpha (NR1F1) did not. Transcriptional activities of the receptors whose DNA binding domains (DBDs) were replaced by the GAL4 DBD were repressed by Dax-1 to various levels, which correlated with the strength of interaction. Amino acid substitutions revealed that Ad4BP/SF-1 and LRH-1 preferentially interact with L(+1)XXLL-related motifs containing serine, tyrosine, serine, and threonine at positions -2, +2, +3, and +6, respectively. Taken together, our results indicate that the specificities of LXXLL-related motifs in Dax-1 based on their amino acid sequences play an important role in regulation of orphan receptors.

Citing Articles

Liver receptor homolog-1: structures, related diseases, and drug discovery.

Wu T, Lu Z, Yu H, Wu X, Liu Y, Xu Y Acta Pharmacol Sin. 2024; 45(8):1571-1581.

PMID: 38632319 PMC: 11272790. DOI: 10.1038/s41401-024-01276-x.

A novel variant of NR5A1, p.R350W implicates potential interactions with unknown co-factors or ligands.

Gau M, Suga R, Hijikata A, Kashimada A, Takagi M, Nakagawa R Front Endocrinol (Lausanne). 2023; 13:1033074.

PMID: 36743925 PMC: 9895113. DOI: 10.3389/fendo.2022.1033074.

Nuclear receptor subfamily 5 group A member 2 (NR5A2): role in health and diseases.

Sandhu N, Rana S, Meena K Mol Biol Rep. 2021; 48(12):8155-8170.

PMID: 34643922 DOI: 10.1007/s11033-021-06784-1.

Nonsense variant of NR0B1 causes hormone disorders associated with congenital adrenal hyperplasia.

Fan D, Li L, Zhang H Sci Rep. 2021; 11(1):16066.

PMID: 34373561 PMC: 8352982. DOI: 10.1038/s41598-021-95642-y.

C2H2-Type Zinc Finger Proteins: Evolutionarily Old and New Partners of the Nuclear Hormone Receptors.

Mackeh R, Marr A, Fadda A, Kino T Nucl Recept Signal. 2019; 15:1550762918801071.

PMID: 30718982 PMC: 6348741. DOI: 10.1177/1550762918801071.

References

. A unified nomenclature system for the nuclear receptor superfamily. Cell. 1999; 97(2):161-3. DOI: 10.1016/s0092-8674(00)80726-6. View

Nolte R, Wisely G, Westin S, Cobb J, Lambert M, Kurokawa R . Ligand binding and co-activator assembly of the peroxisome proliferator-activated receptor-gamma. Nature. 1998; 395(6698):137-43. DOI: 10.1038/25931. View

James P, Halladay J, Craig E . Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics. 1996; 144(4):1425-36. PMC: 1207695. DOI: 10.1093/genetics/144.4.1425. View

Boerboom D, Pilon N, Behdjani R, Silversides D, Sirois J . Expression and regulation of transcripts encoding two members of the NR5A nuclear receptor subfamily of orphan nuclear receptors, steroidogenic factor-1 and NR5A2, in equine ovarian cells during the ovulatory process. Endocrinology. 2000; 141(12):4647-56. DOI: 10.1210/endo.141.12.7808. View

Lala D, Syka P, Lazarchik S, Mangelsdorf D, Parker K, HEYMAN R . Activation of the orphan nuclear receptor steroidogenic factor 1 by oxysterols. Proc Natl Acad Sci U S A. 1997; 94(10):4895-900. PMC: 24602. DOI: 10.1073/pnas.94.10.4895. View

Egea P, Mitschler A, Rochel N, Ruff M, Chambon P, Moras D . Crystal structure of the human RXRalpha ligand-binding domain bound to its natural ligand: 9-cis retinoic acid. EMBO J. 2000; 19(11):2592-601. PMC: 212755. DOI: 10.1093/emboj/19.11.2592. View

Yussa M, Lohr U, Su K, Pick L . The nuclear receptor Ftz-F1 and homeodomain protein Ftz interact through evolutionarily conserved protein domains. Mech Dev. 2001; 107(1-2):39-53. DOI: 10.1016/s0925-4773(01)00448-8. View

Nachtigal M, Hirokawa Y, Flanagan J, Hammer G, Ingraham H . Wilms' tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex-specific gene expression. Cell. 1998; 93(3):445-54. DOI: 10.1016/s0092-8674(00)81172-1. View

Voegel J, Heine M, Zechel C, Chambon P, Gronemeyer H . TIF2, a 160 kDa transcriptional mediator for the ligand-dependent activation function AF-2 of nuclear receptors. EMBO J. 1996; 15(14):3667-75. PMC: 452006. View

10.

Altincicek B, Tenbaum S, Dressel U, Thormeyer D, Renkawitz R, Baniahmad A . Interaction of the corepressor Alien with DAX-1 is abrogated by mutations of DAX-1 involved in adrenal hypoplasia congenita. J Biol Chem. 2000; 275(11):7662-7. DOI: 10.1074/jbc.275.11.7662. View

11.

Heery D, Hoare S, Hussain S, Parker M, SHEPPARD H . Core LXXLL motif sequences in CREB-binding protein, SRC1, and RIP140 define affinity and selectivity for steroid and retinoid receptors. J Biol Chem. 2000; 276(9):6695-702. DOI: 10.1074/jbc.M009404200. View

12.

Warnmark A, Almlof T, Leers J, Gustafsson J, Treuter E . Differential recruitment of the mammalian mediator subunit TRAP220 by estrogen receptors ERalpha and ERbeta. J Biol Chem. 2001; 276(26):23397-404. DOI: 10.1074/jbc.M011651200. View

13.

Kane C, Means A . Activation of orphan receptor-mediated transcription by Ca(2+)/calmodulin-dependent protein kinase IV. EMBO J. 2000; 19(4):691-701. PMC: 305607. DOI: 10.1093/emboj/19.4.691. View

14.

Ko L, Cardona G, Iwasaki T, Bramlett K, Burris T, Chin W . Ser-884 adjacent to the LXXLL motif of coactivator TRBP defines selectivity for ERs and TRs. Mol Endocrinol. 2002; 16(1):128-40. DOI: 10.1210/mend.16.1.0755. View

15.

Lee Y, Dell H, Dowhan D, Moore D . The orphan nuclear receptor SHP inhibits hepatocyte nuclear factor 4 and retinoid X receptor transactivation: two mechanisms for repression. Mol Cell Biol. 1999; 20(1):187-95. PMC: 85074. DOI: 10.1128/MCB.20.1.187-195.2000. View

16.

Luo X, Ikeda Y, Parker K . A cell-specific nuclear receptor is essential for adrenal and gonadal development and sexual differentiation. Cell. 1994; 77(4):481-90. DOI: 10.1016/0092-8674(94)90211-9. View

17.

Ikeda Y, Takeda Y, Shikayama T, Mukai T, Hisano S, Morohashi K . Comparative localization of Dax-1 and Ad4BP/SF-1 during development of the hypothalamic-pituitary-gonadal axis suggests their closely related and distinct functions. Dev Dyn. 2001; 220(4):363-76. DOI: 10.1002/dvdy.1116. View

18.

Shinoda K, Lei H, Yoshii H, Nomura M, Nagano M, Shiba H . Developmental defects of the ventromedial hypothalamic nucleus and pituitary gonadotroph in the Ftz-F1 disrupted mice. Dev Dyn. 1995; 204(1):22-9. DOI: 10.1002/aja.1002040104. View

19.

Ito M, Yu R, Jameson J . Steroidogenic factor-1 contains a carboxy-terminal transcriptional activation domain that interacts with steroid receptor coactivator-1. Mol Endocrinol. 1998; 12(2):290-301. DOI: 10.1210/mend.12.2.0059. View

20.

Forman B, Umesono K, Chen J, Evans R . Unique response pathways are established by allosteric interactions among nuclear hormone receptors. Cell. 1995; 81(4):541-50. DOI: 10.1016/0092-8674(95)90075-6. View