title: Dissecting glucose signalling with diversity-oriented synthesis and small-molecule microarrays.
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.
authors.affiliation: Howard Hughes Medical Institute, Institute for Chemistry and Cell Biology, Bauer Center for Genomics Research, Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: Finny G
authors.initials: FG
authors.lastname: Kuruvilla
authors.forename: Alykhan F
authors.initials: AF
authors.lastname: Shamji
authors.forename: Scott M
authors.initials: SM
authors.lastname: Sternson
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hergenrother
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Schreiber
mesh_terms: D004146:Dioxanes / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005947:Glucose / Q000378:metabolism*; D005979:Glutathione Peroxidase; D008024:Ligands; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D010080:Oxazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics / Q000494:pharmacology; D011328:Prions; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D004146:Dioxanes; D008024:Ligands; D010080:Oxazoles; D011328:Prions; D012333:RNA, Messenger; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C454543:uretupamine A; C454542:uretupamine B; D005979:Glutathione Peroxidase; C067020:URE2 protein, S cerevisiae; D005947:Glucose