Abstract
Small molecules that alter protein function provide a means to modulate biological networks with temporal resolution. Here we demonstrate a potentially general and scalable method of identifying such molecules by application to a particular protein, Ure2p, which represses the transcription factors Gln3p and Nil1p. By probing a high-density microarray of small molecules generated by diversity-oriented synthesis with fluorescently labelled Ure2p, we performed 3,780 protein-binding assays in parallel and identified several compounds that bind Ure2p. One compound, which we call uretupamine, specifically activates a glucose-sensitive transcriptional pathway downstream of Ure2p. Whole-genome transcription profiling and chemical epistasis demonstrate the remarkable Ure2p specificity of uretupamine and its ability to modulate the glucose-sensitive subset of genes downstream of Ure2p. These results demonstrate that diversity-oriented synthesis and small-molecule microarrays can be used to identify small molecules that bind to a protein of interest, and that these small molecules can regulate specific functions of the protein.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Dioxanes / chemical synthesis
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Dioxanes / chemistry
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Dioxanes / metabolism*
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Dioxanes / pharmacokinetics
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Dioxanes / pharmacology
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Dose-Response Relationship, Drug
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Gene Expression Profiling
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Gene Expression Regulation, Fungal* / drug effects
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Glucose / metabolism*
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Glutathione Peroxidase
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Ligands
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Models, Biological
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Oligonucleotide Array Sequence Analysis
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Oxazoles / chemical synthesis
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Oxazoles / chemistry
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Oxazoles / metabolism*
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Oxazoles / pharmacokinetics
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Oxazoles / pharmacology
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Prions*
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Protein Binding
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Repressor Proteins / agonists
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Repressor Proteins / antagonists & inhibitors
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Repressor Proteins / metabolism
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Saccharomyces cerevisiae / drug effects
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / agonists
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Saccharomyces cerevisiae Proteins / antagonists & inhibitors
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Signal Transduction* / drug effects
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Structure-Activity Relationship
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Substrate Specificity
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Transcription, Genetic / drug effects
Substances
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Dioxanes
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Ligands
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Oxazoles
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Prions
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RNA, Messenger
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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uretupamine A
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uretupamine B
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Glutathione Peroxidase
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URE2 protein, S cerevisiae
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Glucose