Abstract
The human nuclear receptor liver receptor homolog 1 (hLRH-1) plays an important role in the development of breast carcinomas. This orphan receptor is efficiently downregulated by the unusual co-repressor SHP and has been thought to be ligand-independent. We present the crystal structure at a resolution of 1.9 Å of the ligand-binding domain of hLRH-1 in complex with the NR box 1 motif of human SHP, which we find contacts the AF-2 region of hLRH-1 using selective structural motifs. Electron density indicates phospholipid bound within the ligand-binding pocket, which we confirm using mass spectrometry of solvent-extracted samples. We further show that pocket mutations reduce phospholipid binding and receptor activity in vivo. Our results indicate that hLRH-1's control of gene expression is mediated by phospholipid binding, and establish hLRH-1 as a novel target for compounds designed to slow breast cancer development.
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06 March 2005
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Notes
Note: In the version of this article originally published online, Figure 4c and the legend describing it contained mistakes. The second and third lines in the graphical legends for this panel read “SRC-3” and “SRC-2”, respectively. The text read “ (c) The F342W I416W form of hLRH-1 LBD, which is reduced in phospholipid binding, shows reduced levels of coactivation by SRC-2 and SRC-3.” We apologize for any inconvenience this may have caused. This error has been corrected for the HTML and print versions of the article.
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Acknowledgements
We thank S. Lujan and G. Pielak for discussions regarding biophysical techniques; D. Fargo (UNC Center for Bioinformatics) for assistance in promoter sequence searching; and members of the Redinbo laboratory for experimental assistance and stimulating discussions. This work was supported by US National Institutes of Health grant DK62229 (M.R.R.). Nondenaturing mass spectrometry analysis was partially supported by an anonymous gift to the UNC Michael Hooker Proteomics Core Facility for research targeted to proteomics and cystic fibrosis, and was conducted by core staff members C. Scarlett and V. Mocanu. E.A.O. was supported by a Lineberger Comprehensive Cancer Center postdoctoral fellowship, and I.H.S. and J.M.H. were supported in part by the UNC Smallwood Foundation and Office of Undergraduate Research. The mass spectrometry facility in the Department of Biochemistry of the Duke University Medical Center and Z.G. were supported by Core K of LIPID MAPS consortium grant 1U54GM069338.
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Ortlund, E., Lee, Y., Solomon, I. et al. Modulation of human nuclear receptor LRH-1 activity by phospholipids and SHP. Nat Struct Mol Biol 12, 357–363 (2005). https://doi.org/10.1038/nsmb910
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DOI: https://doi.org/10.1038/nsmb910
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