Research Summary

My lab is focused on structure function studies of nuclear receptors and transcription control proteins in stem cells. My lab uses biochemical and biophysical methods to evaluate protein complexes. Among the technologies employed are surface plasmon resonance, X-ray crystallography, and differential scanning fluorescence. Mutagenesis and cell assays are employed to gauge functional effects of mutagenesis to evaluate the hypotheses linking structure to function. We determined the first three-dimensional structure for any nuclear receptor and published papers on the thyroid hormone receptor, androgen receptor, and glucocorticoid receptor, as well as on LRH-1, SF-1, and Dax-1 receptors, and developed bait-affinity expression for making the first complex of a nuclear receptor with its corepressor. Research on androgen receptor interactions with peptides and small molecules helped define the specificity of the AF2 site and led to the discovery of a new protein binding site on the androgen receptor hormone binding domain.

My lab is known for determining structures of important protein molecules and assemblies, and relating structures to the mechanisms of function of the macromolecules. Many of the papers from the lab are recognized by experts as useful contributions to the field. Early papers from my lab were mostly on learning about allosteric control of enzyme catalysis. Using glycogen phosphorylase as the target, we determined the structure at high resolution for the phosphorylated dimer, normally active but inhibited by an allosteric effector. At about 200,000 molecular weight, this protein was the largest with known structure for about ten years. We also embraced recombinant DNA methods early. Ours was the first lab to clone all isozymes for any human protein. Our muscle cDNA library was used by others to clone challenging targets, including the first Ca ATPase. With expertise in cloning technologies, we were positioned to compare the regulation of glycogen phosphorylases in evolution from yeast to human. Recognition of my work on allostery was the basis for election to the National Academy of Sciences. Recently, our lab has focused on learning more about mechanisms of transcription factors, especially nuclear receptors. This focus led us to publish our first paper in cancer biology that identifies a potential target for treating pancreatic cancer.

Research Funding

  • September 30, 2010 - June 30, 2015 - Structures of Protein Complexes Regulating Transcription in Enbryonic Stem Cells, Principal Investigator. Sponsor: NIH/NIGMS, Sponsor Award ID: U01GM094614
  • March 1, 1980 - February 28, 2015 - A Synchroton Radiation Structural Biology Resource, Co-Investigator. Sponsor: NIH, Sponsor Award ID: P41RR001209
  • January 15, 2012 - December 31, 2013 - Screening for antagonists of nuclear receptor LRH-1 in pancreatic cancer cells, Principal Investigator. Sponsor: NIH/NIMH, Sponsor Award ID: R03MH094165
  • September 1, 1983 - June 30, 2013 - Macromolecular Diffraction Resource: MacCHESS, Co-Investigator. Sponsor: NIH, Sponsor Award ID: P41RR001646

Education

Marietta College, Marietta, Ohio, B.S., 1965, Chemistry
Cornell University, Ithaca, New York, Ph.D., 1970, Physical Chemistry
Yale University, New Haven, Connecticut, Department of Molecular Biophysics and Biochemistry, Laboratory of Prof. Thomas Steitz, Postdoctoral Fellow, 1970-1971, X-ray Crystallography
Yale University, New Haven, Connecticut, Department of Biophysics and Biochemistry, Postdoctoral Fellow, 1971-1973, X-ray Crystallography

Honors & Awards

  • 1965
    Graduated with Honors in Chemistry, Hobba Chemistry Prize
  • 1965-1970
    National Institutes of Health Research Studentship
  • 1970-1971
    National Institutes of Health Postdoctoral Fellowship
  • 1971-1973
    National Institutes of Health Postdoctoral Fellowship
  • 1983
    Robert Welch Foundation Lecturer
  • Federal and Public Scientific Advisory Boards
  • 1991
    Member, Structural Biology Committee, Federal Government of Spain
  • 1993-1995
    Member, Standing Committee, Protein Engineering Network, Medical Research Council of Canada
  • 1993-1994
    Structural Biology Scientific Advisory Board, Institute of Molecular Biology, Academia Sinica, Nankang, Taipei
  • 1993-1997
    Member, Scientific Advisory Committee of the Cancer Research Fund, Damon Runyon-Walter Winchell Foundation
  • 1994-present
    Protein Society Nominating Committee
  • 1997-2000
    University of California Biotechnology Star Program

Selected Publications

  1. Bruning JM, Wang Y, Oltrabella F, Tian B, Kholodar SA, Liu H, Bhattacharya P, Guo S, Holton JM, Fletterick RJ, Jacobson MP, England PM. Covalent Modification and Regulation of the Nuclear Receptor Nurr1 by a Dopamine Metabolite. Cell Chem Biol. 2019 05 16; 26(5):674-685.e6.  View on PubMed
  2. Bayrer JR, Wang H, Nattiv R, Suzawa M, Escusa HS, Fletterick RJ, Klein OD, Moore DD, Ingraham HA. LRH-1 mitigates intestinal inflammatory disease by maintaining epithelial homeostasis and cell survival. Nat Commun. 2018 10 10; 9(1):4055.  View on PubMed
  3. de Jesus Cortez F, Nguyen P, Truillet C, Tian B, Kuchenbecker KM, Evans MJ, Webb P, Jacobson MP, Fletterick RJ, England PM. Development of 5N-Bicalutamide, a High-Affinity Reversible Covalent Antiandrogen. ACS Chem Biol. 2017 12 15; 12(12):2934-2939.  View on PubMed
  4. Fletterick R. NR5A2 discovering compounds that block tumor growth in PDAC. J Surg Oncol. 2017 Jul; 116(1):89-93.  View on PubMed
  5. de Jesus Cortez F, Suzawa M, Irvy S, Bruning JM, Sablin E, Jacobson MP, Fletterick RJ, Ingraham HA, England PM. Disulfide-Trapping Identifies a New, Effective Chemical Probe for Activating the Nuclear Receptor Human LRH-1 (NR5A2). PLoS One. 2016; 11(7):e0159316.  View on PubMed
  6. Proudfoot A, Axelrod HL, Geralt M, Fletterick RJ, Yumoto F, Deacon AM, Elsliger MA, Wilson IA, Wüthrich K, Serrano P. Dlx5 Homeodomain:DNA Complex: Structure, Binding and Effect of Mutations Related to Split Hand and Foot Malformation Syndrome. J Mol Biol. 2016 Mar 27; 428(6):1130-1141.  View on PubMed
  7. Suzawa M, Miranda DA, Ramos KA, Ang KK, Faivre EJ, Wilson CG, Caboni L, Arkin MR, Kim YS, Fletterick RJ, Diaz A, Schneekloth JS, Ingraham HA. A gene-expression screen identifies a non-toxic sumoylation inhibitor that mimics SUMO-less human LRH-1 in liver. Elife. 2015 Dec 11; 4.  View on PubMed
  8. Sablin EP, Blind RD, Uthayaruban R, Chiu HJ, Deacon AM, Das D, Ingraham HA, Fletterick RJ. Structure of Liver Receptor Homolog-1 (NR5A2) with PIP3 hormone bound in the ligand binding pocket. J Struct Biol. 2015 Dec; 192(3):342-348.  View on PubMed
  9. Storer Samaniego C, Suh JH, Chattopadhyay A, Olivares K, Guy N, Sivils JC, Dey P, Yumoto F, Fletterick RJ, Strom AM, Gustafsson JÅ, Webb P, Cox MB. The FKBP52 Cochaperone Acts in Synergy with ß-Catenin to Potentiate Androgen Receptor Signaling. PLoS One. 2015; 10(7):e0134015.  View on PubMed
  10. Hayashi Y, Caboni L, Das D, Yumoto F, Clayton T, Deller MC, Nguyen P, Farr CL, Chiu HJ, Miller MD, Elsliger MA, Deacon AM, Godzik A, Lesley SA, Tomoda K, Conklin BR, Wilson IA, Yamanaka S, Fletterick RJ. Structure-based discovery of NANOG variant with enhanced properties to promote self-renewal and reprogramming of pluripotent stem cells. Proc Natl Acad Sci U S A. 2015 Apr 14; 112(15):4666-71.  View on PubMed
  11. Torres IO, Kuchenbecker KM, Nnadi CI, Fletterick RJ, Kelly MJ, Fujimori DG. Histone demethylase KDM5A is regulated by its reader domain through a positive-feedback mechanism. Nat Commun. 2015 Feb 17; 6:6204.  View on PubMed
  12. Bayrer JR, Mukkamala S, Sablin EP, Webb P, Fletterick RJ. Silencing LRH-1 in colon cancer cell lines impairs proliferation and alters gene expression programs. Proc Natl Acad Sci U S A. 2015 Feb 24; 112(8):2467-72.  View on PubMed
  13. Gallastegui N, Mackinnon JA, Fletterick RJ, Estébanez-Perpiñá E. Advances in our structural understanding of orphan nuclear receptors. Trends Biochem Sci. 2015 Jan; 40(1):25-35.  View on PubMed
  14. Blind RD, Sablin EP, Kuchenbecker KM, Chiu HJ, Deacon AM, Das D, Fletterick RJ, Ingraham HA. The signaling phospholipid PIP3 creates a new interaction surface on the nuclear receptor SF-1. Proc Natl Acad Sci U S A. 2014 Oct 21; 111(42):15054-9.  View on PubMed
  15. Nagavalli S, Fletterick RJ, Ford G, Boston BA, Gianoukakis AG. Computer modeling of a newly identified THRB gene mutation (S350L) associated with resistance to thyroid hormone in three unrelated patients. Thyroid. 2014 Sep; 24(9):1430-1.  View on PubMed
  16. Benod C, Villagomez R, Filgueira CS, Hwang PK, Leonard PG, Poncet-Montange G, Rajagopalan S, Fletterick RJ, Gustafsson JÅ, Webb P. The human orphan nuclear receptor tailless (TLX, NR2E1) is druggable. PLoS One. 2014; 9(6):e99440.  View on PubMed
  17. Bhattacharya S, Lou X, Hwang P, Rajashankar KR, Wang X, Gustafsson JÅ, Fletterick RJ, Jacobson RH, Webb P. Structural and functional insight into TAF1-TAF7, a subcomplex of transcription factor II D. Proc Natl Acad Sci U S A. 2014 Jun 24; 111(25):9103-8.  View on PubMed
  18. Larsen CC, Dumitrescu A, Guerra-Argüero LM, Gállego-Suárez C, Vazquez-Mellado A, Vinogradova M, Fletterick R, Refetoff S, Weiss RE. Incidental identification of a thyroid hormone receptor beta (THRB) gene variant in a family with autoimmune thyroid disease. Thyroid. 2013 Dec; 23(12):1638-43.  View on PubMed
  19. Vinogradova MV, Malanina GG, Waitzman JS, Rice SE, Fletterick RJ. Plant Kinesin-Like Calmodulin Binding Protein Employs Its Regulatory Domain for Dimerization. PLoS One. 2013; 8(6):e66669.  View on PubMed
  20. Benod C, Carlsson J, Uthayaruban R, Hwang P, Irwin JJ, Doak AK, Shoichet BK, Sablin EP, Fletterick RJ. Structure-based discovery of antagonists of nuclear receptor LRH-1. J Biol Chem. 2013 Jul 05; 288(27):19830-44.  View on PubMed

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