TY - JOUR
T1 - Dissecting glucose signalling with diversity-oriented synthesis and small-molecule microarrays
AU - Kuruvilla, Finny G.
AU - Shamji, Alykhan F.
AU - Sternson, Scott M.
AU - Hergenrother, Paul J.
AU - Schreiber, Stuart L.
N1 - Funding Information:
We thank R. Melki for providing bacterially expressed Ure2p protein. F.G.K. was supported by the NIH Medical Scientist Training Program, A.F.S. by the Howard Hughes Medical Institute predoctoral fellowship, S.M.S. by the Roche and NSF predoctoral fellowships, and P.J.H. by the American Cancer Society. S.L.S. is an Investigator at the Howard Hughes Medical Institute. This research was funded by a grant from the NIGMS (GM-38627).
Funding Information:
We thank R. Baer, R. Dalla-Favera, B. Tycko and T. Ludwig for critical discussions; we also thank many colleges in the field for providing antibodies, cell lines and plasmids, and other members of W.G.’s laboratory for sharing unpublished data and critical comments. This work was supported in part by grants from Avon Foundation, the Stewart Trust, the Irma T. Hirschl Trust and NIH/NCI to W.G., who is also a Leukemia and Lymphoma Society Scholar.
PY - 2002/4/11
Y1 - 2002/4/11
N2 - 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.
AB - 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.
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U2 - 10.1038/416653a
DO - 10.1038/416653a
M3 - Article
C2 - 11948353
AN - SCOPUS:0037061492
SN - 0028-0836
VL - 416
SP - 653
EP - 657
JO - Nature
JF - Nature
IS - 6881
ER -