title: A gene-expression screen identifies a non-toxic sumoylation inhibitor that mimics SUMO-less human LRH-1 in liver.
abstract: SUMO-modification of nuclear proteins has profound effects on gene expression. However, non-toxic chemical tools that modulate sumoylation in cells are lacking. Here, to identify small molecule sumoylation inhibitors we developed a cell-based screen that focused on the well-sumoylated substrate, human Liver Receptor Homolog-1 (hLRH-1, NR5A2). Our primary gene-expression screen assayed two SUMO-sensitive transcripts, APOC3 and MUC1, that are upregulated by SUMO-less hLRH-1 or by siUBC9 knockdown, respectively. A polyphenol, tannic acid (TA) emerged as a potent sumoylation inhibitor in vitro (IC50 = 12.8 µM) and in cells. TA also increased hLRH-1 occupancy on SUMO-sensitive transcripts. Most significantly, when tested in humanized mouse primary hepatocytes, TA inhibits hLRH-1 sumoylation and induces SUMO-sensitive genes, thereby recapitulating the effects of expressing SUMO-less hLRH-1 in mouse liver. Our findings underscore the benefits of phenotypic screening for targeting post-translational modifications, and illustrate the potential utility of TA for probing the cellular consequences of sumoylation.
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
authors.forename: Miyuki
authors.initials: M
authors.lastname: Suzawa
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
authors.forename: Diego A
authors.initials: DA
authors.lastname: Miranda
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
authors.forename: Karmela A
authors.initials: KA
authors.lastname: Ramos
authors.affiliation: Small Molecule Discovery Center, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States.
authors.forename: Kenny K-H
authors.initials: KK
authors.lastname: Ang
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Faivre
authors.affiliation: Small Molecule Discovery Center, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Wilson
authors.affiliation: Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States.
authors.forename: Laura
authors.initials: L
authors.lastname: Caboni
authors.affiliation: Small Molecule Discovery Center, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States.
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Arkin
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute, Frederick, United States.
authors.forename: Yeong-Sang
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Fletterick
authors.affiliation: Department of Epidemiology and Biostatistics, University of California, San Francisco, San Francisco, United States.
authors.forename: Aaron
authors.initials: A
authors.lastname: Diaz
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute, Frederick, United States.
authors.forename: John S
authors.initials: JS
authors.lastname: Schneekloth
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States.
authors.forename: Holly A
authors.initials: HA
authors.lastname: Ingraham
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D016513:Mice, SCID; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D058207:Sumoylation / Q000187:drug effects*; D013634:Tannins / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; C082631:NR5A2 protein, human; D018160:Receptors, Cytoplasmic and Nuclear; D013634:Tannins
keywords: NASH; NR5As; cell biology; human; human biology; medicine; mouse; phenotypic screen; primary hepatocytes; sumoylation; tannic acid