A gene-expression screen identifies a non-toxic sumoylation inhibitor that mimics SUMO-less human LRH-1 in liver

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, hum...

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Bibliographic Details
Published in:	eLife Vol. 4
Main Authors:	Suzawa, Miyuki, Miranda, Diego A, Ramos, Karmela A, Ang, Kenny K-H, Faivre, Emily J, Wilson, Christopher G, Caboni, Laura, Arkin, Michelle R, Kim, Yeong-Sang, Fletterick, Robert J, Diaz, Aaron, Schneekloth, John S, Ingraham, Holly A
Format:	Journal Article
Language:	English
Published:	England eLife Science Publications, Ltd 11-12-2015 eLife Sciences Publications Ltd eLife Sciences Publications, Ltd
Subjects:	Animals Cell Biology Cells, Cultured Drug Evaluation, Preclinical - methods Enzyme Inhibitors - isolation & purification Enzyme Inhibitors - metabolism Gene expression Gene Expression Profiling Genetic aspects Hepatocytes - drug effects Hepatocytes - enzymology Human Biology and Medicine Humans Inhibitory Concentration 50 Liver Mice Mice, SCID NASH NR5As Observations phenotypic screen primary hepatocytes Receptors, Cytoplasmic and Nuclear - metabolism sumoylation Sumoylation - drug effects tannic acid Tannins - isolation & purification Tannins - metabolism Tools and Resources human biology mouse NR5As cell biology sumoylation primary hepatocytes phenotypic screen medicine tannic acid NASH human
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Summary:	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.
Bibliography:	These authors contributed equally to this work.
ISSN:	2050-084X 2050-084X
DOI:	10.7554/eLife.09003