title: A rapid and sensitive high-throughput screening method to identify compounds targeting protein-nucleic acids interactions.
abstract: DNA-binding and RNA-binding proteins are usually considered 'undruggable' partly due to the lack of an efficient method to identify inhibitors from existing small molecule repositories. Here we report a rapid and sensitive high-throughput screening approach to identify compounds targeting protein-nucleic acids interactions based on protein-DNA or protein-RNA interaction enzyme-linked immunosorbent assays (PDI-ELISA or PRI-ELISA). We validated the PDI-ELISA method using the mammalian high-mobility-group protein AT-hook 2 (HMGA2) as the protein of interest and netropsin as the inhibitor of HMGA2-DNA interactions. With this method we successfully identified several inhibitors and an activator for HMGA2-DNA interactions from a collection of 29 DNA-binding compounds. Guided by this screening excise, we showed that netropsin, the specific inhibitor of HMGA2-DNA interactions, strongly inhibited the differentiation of the mouse pre-adipocyte 3T3-L1 cells into adipocytes, most likely through a mechanism by which the inhibition is through preventing the binding of HMGA2 to the target DNA sequences. This method should be broadly applicable to identify compounds or proteins modulating many DNA-binding or RNA-binding proteins.
authors: Biomolecular Sciences Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA Department of Chemistry & Biochemistry, Florida International University, 11200 SW 8th Street, FL 33199, USA.
authors: Nicole
authors: N
authors: Alonso
authors: Biomolecular Sciences Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA Department of Chemistry & Biochemistry, Florida International University, 11200 SW 8th Street, FL 33199, USA.
authors: Roboan
authors: R
authors: Guillen
authors: Biomolecular Sciences Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA Department of Cellular Biology and Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL 33199, USA.
authors: Jeremy W
authors: JW
authors: Chambers
authors: Biomolecular Sciences Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA Department of Chemistry & Biochemistry, Florida International University, 11200 SW 8th Street, FL 33199, USA lengf@fiu.edu.
authors: Fenfei
authors: F
authors: Leng
mesh_terms: D041721:3T3-L1 Cells; D050156:Adipogenesis / Q000187:drug effects; D000818:Animals; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D025743:HMGA2 Protein / Q000037:antagonists & inhibitors / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D051379:Mice; D009429:Netropsin / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D025743:HMGA2 Protein; D016601:RNA-Binding Proteins; D009429:Netropsin; D004247:DNA
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