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Antidiabetic phospholipid–nuclear receptor complex reveals the mechanism for phospholipid-driven gene regulation

Nature Structural & Molecular Biology volume 19, pages 532–537 (2012)Cite this article

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Abstract

The human nuclear receptor liver receptor homolog-1 (LRH-1) has an important role in controlling lipid and cholesterol homeostasis and is a potential target for the treatment of diabetes and hepatic diseases. LRH-1 is known to bind phospholipids, but the role of phospholipids in controlling LRH-1 activation remains highly debated. Here we describe the structure of both apo LRH-1 and LRH-1 in complex with the antidiabetic phospholipid dilauroylphosphatidylcholine (DLPC). Together with hydrogen-deuterium exchange MS and functional data, our studies show that DLPC binding is a dynamic process that alters co-regulator selectivity. We show that the lipid-free receptor undergoes previously unrecognized structural fluctuations, allowing it to interact with widely expressed co-repressors. These observations enhance our understanding of LRH-1 regulation and highlight its importance as a new therapeutic target for controlling diabetes.

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Figure 1: DLPC binds directly to LRH-1 and promotes activation through unique interactions.
Figure 2: DLPC alters LRH-1 stability and structural dynamics.
Figure 3: DLPC binding affects LRH-1 dynamics and co-regulator preference.
Figure 4: Structure of apo LRH-1 identifies a novel functional region for activation.

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Acknowledgements

We thank N.T. Seyfried in the Department of Biochemistry at Emory University for his help in acquiring the MS data for the proteolysis protection assays. We thank F.H. Strobel in the Department of Chemistry at Emory University for his help in acquiring the MS data for phospholipids. P.R.G. was supported by NIH grants GM084041 (PI: P.R. Griffin) and MH084512 (PI: H. Rosen). This work was supported with start-up funds from Emory University. P.M.M. was supported by an Emory–National Institute of Environmental Health Sciences Graduate and Postdoctoral Training in Toxicology grant (T32ES012870).

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Author notes

  1. Paul M Musille and Manish C Pathak: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry, Winship Cancer Institute, Emory University School of Medicine, Atlanta, Georgia, USA

    Paul M Musille, Manish C Pathak, William H Hudson & Eric A Ortlund

  2. Discovery and Developmental Therapeutics, Winship Cancer Institute, Emory University School of Medicine, Atlanta, Georgia, USA

    Paul M Musille, Manish C Pathak, William H Hudson & Eric A Ortlund

  3. Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida, USA

    Janelle L Lauer & Patrick R Griffin

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Contributions

P.M.M. purified, crystallized and determined both LRH-1 structures, measured phospholipid levels and carried out co-regulator-peptide interaction studies. M.C.P. measured lipid phosphorus levels, optimized apo LRH-1 production and carried out co-regulator peptide interaction studies. W.H.H. conducted reporter gene experiments. J.L.L. and P.R.G. conducted thermal unfolding and HDX experiments. P.M.M., J.L.L., P.R.G. and E.A.O. analyzed and interpreted the data. P.M.M. and E.A.O. conceived the experiments and wrote the manuscript.

Corresponding author

Correspondence to Eric A Ortlund.

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The authors declare no competing financial interests.

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Supplementary Figures 1–4, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 10797 kb)

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Musille, P., Pathak, M., Lauer, J. et al. Antidiabetic phospholipid–nuclear receptor complex reveals the mechanism for phospholipid-driven gene regulation. Nat Struct Mol Biol 19, 532–537 (2012). https://doi.org/10.1038/nsmb.2279

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