Scientific Publications

Discovery of an inhibitor of a transcription factor using small molecule microarrays and diversity-oriented synthesis.

Publication TypeJournal Article
AuthorsKoehler, AN, Shamji AF, and Schreiber SL
AbstractSmall molecule microarrays were screened to identify a small molecule ligand for Hap3p, a subunit of the yeast Hap2/3/4/5p transcription factor complex. The compound, named haptamide A, was determined to have a KD of 5.03 muM for binding to Hap3p using surface plasmon resonance analysis. Haptamide A also inhibited activation of a GDH1-lacZ reporter gene in a dose-dependent fashion. To explore structure-activity relationships, 11 derivatives of haptamide A were prepared using the same synthetic route that was developed for the original library synthesis. Analysis of dissociation constants and IC50 values for the reporter gene assay revealed a more potent inhibitor, haptamide B, with a KD of 330 nM. Whole-genome transcriptional profiling was used to compare effects of haptamide B with a hap3Delta yeast strain. Treatment with haptamide B, like the deletion mutant, reduced lactate-induced transcription of several genes from wild-type levels. Profiling the genetic "knockout" and the chemical genetic "knockdown" led to the identification of several genes that are regulated by Hap3p under nonfermentative conditions. These results demonstrate that a small molecule discovered using the small molecule microarray binding assay can permeate yeast cells and reach its target transcription factor protein in cells.
Year of Publication2003
JournalJournal of the American Chemical Society
Volume125
Issue28
Pages8420-1
Date Published (YYYY/MM/DD)2003/07/16
ISSN Number0002-7863
DOI10.1021/ja0352698
PubMedhttp://www.ncbi.nlm.nih.gov/pubmed/12848532?dopt=Abstract