Principal Investigator/Program Director (Last, first, middle): Fletterick, Robert, J. RESEARCH &RELATED Other Project Information 1. * Are Human Subjects Involved? m Yes l No 1.a. If YES to Human Subjects Is the IRB review Pending? m Yes m No IRB Approval Date: Exemption Number: 1 2 3 4 5 6 Human Subject Assurance Number 2. * Are Vertebrate Animals Used? m Yes l No 2.a. If YES to Vertebrate Animals Is the IACUC review Pending? m Yes m No IACUC Approval Date: Animal Welfare Assurance Number 3. * Is proprietary/privileged information m Yes l No included in the application? 4.a.* Does this project have an actual or potential impact on m Yes l No the environment? 4.b. If yes, please explain: 4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an environmental assessment (EA) or environmental impact statement (EIS) been performed? m Yes m No 4.d. If yes, please explain: 5.a.* Does this project involve activities outside the U.S. or m Yes l No partnership with International Collaborators? 5.b. If yes, identify countries: 5.c. Optional Explanation: 6. * Project Summary/Abstract Abstract1001854800.pdf Mime Type: application/pdf 7. * Project Narrative ProjectNarrative1001854801.pdf Mime Type: application/pdf 8. Bibliography &References Cited Bibliography1001854802.pdf Mime Type: application/pdf 9. Facilities &Other Resources Facilities1001854804.pdf Mime Type: application/pdf 10. Equipment Equipment1001854805.pdf Mime Type: application/pdf Tracking Number: Other Information Page 5 OMB Number: 4040-0001 Expiration Date: 04/30/2008 Principal Investigator/Program Director (Last, first, middle): Fletterick, Robert, J. Abstract Nuclear receptors program many endocrine processes essential for health and life. Among the most enigmatic are those receptors, which are critically important but have unknown regulating hormones, such as DAX-1, LRH-1 and SF-1. These are the targets of this proposal. Medical disorders teach us about the interplay of these receptors in the endocrine system without telling us their mechanisms. The DAX-1 gene codes an unusual nuclear receptor lacking a DNA interaction domain. Yet it controls the activity of many target genes in many tissues. Mutations in DAX-1 affect embryonic stem cell progression, development of the adrenal and pituitary glands, the hypothalamus and the ovaries or testes leading to severe endocrine disorders. The receptors LRH-1 and SF-1 are critical in developmental processes, and their constitutive activation is muted by DAX-1. These three nuclear receptors and transcription factors are called orphans because their controlling hormones, if they exist, have not been identified. The most basic details of the mechanisms of regulation of these related nuclear receptors are not known. The goal of this proposal is to understand the principles and atomic level details of the mechanisms of regulation of LRH-1 and SF-1 by DAX-1. Since it halts transcriptional activation, DAX-1 may be classed as a co-repressor with unknown hormone. Because regulation by DAX-1 depends on the associations of these proteins we will analyze, at atomic resolution, the binding interactions of DAX-1 with LRH-1 and SF-1. DAX-1 protein, which has never been prepared in a functionally active state for either biochemical or structural studies, will be isolated and crystallized with LRH-1 and SF-1 so that three-dimensional structures of the assemblies may be analyzed. Functional studies guided by the structural models will carried out in cells to reveal the mechanisms of action of these receptors. This information will be the foundation for understanding the molecular pathology of numerous human endocrine syndromes and suggest pathways for therapeutic intervention. The nuclear receptors we are studying are associated with many disease processes, including cancer, infertility, genetic metabolic diseases, and mental health defects. Learning the mechanisms of action of these receptors, and what goes wrong in the diseases, will help suggest pathways for design of new drug treatments for these conditions. Project Description Page 6
| Fletterick, Robert (2017) NR5A2 discovering compounds that block tumor growth in PDAC. J Surg Oncol 116:89-93 |
| Sablin, Elena P; Blind, Raymond D; Uthayaruban, Rubatharshini et al. (2015) Structure of Liver Receptor Homolog-1 (NR5A2) with PIP3 hormone bound in the ligand binding pocket. J Struct Biol 192:342-348 |
| Blind, Raymond D; Sablin, Elena P; Kuchenbecker, Kristopher M et al. (2014) The signaling phospholipid PIP3 creates a new interaction surface on the nuclear receptor SF-1. Proc Natl Acad Sci U S A 111:15054-9 |
| Benod, Cindy; Carlsson, Jens; Uthayaruban, Rubatharshini et al. (2013) Structure-based discovery of antagonists of nuclear receptor LRH-1. J Biol Chem 288:19830-44 |
| Yumoto, Fumiaki; Nguyen, Phuong; Sablin, Elena P et al. (2012) Structural basis of coactivation of liver receptor homolog-1 by ?-catenin. Proc Natl Acad Sci U S A 109:143-8 |
| Benod, Cindy; Vinogradova, Maia V; Jouravel, Natalia et al. (2011) Nuclear receptor liver receptor homologue 1 (LRH-1) regulates pancreatic cancer cell growth and proliferation. Proc Natl Acad Sci U S A 108:16927-31 |
| Sablin, Elena P; Blind, Raymond D; Krylova, Irina N et al. (2009) Structure of SF-1 bound by different phospholipids: evidence for regulatory ligands. Mol Endocrinol 23:25-34 |
| Sablin, Elena P; Woods, April; Krylova, Irina N et al. (2008) The structure of corepressor Dax-1 bound to its target nuclear receptor LRH-1. Proc Natl Acad Sci U S A 105:18390-5 |
