Dissecting glucose signalling with diversity-oriented synthesis and small-molecule microarrays

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Small molecules offer precise control over protein function and biological networks.
• Ure2p protein is a key repressor of transcription factors Gln3p and Nil1p.

Purpose of the Study:

• To develop a general and scalable method for identifying small molecules that modulate protein function.
• To identify compounds that specifically bind and regulate the activity of Ure2p.

Main Methods:

• Utilized diversity-oriented synthesis to create a high-density small molecule microarray.
• Employed fluorescently labeled Ure2p for high-throughput protein-binding assays (3,780 assays).
• Conducted whole-genome transcription profiling and chemical epistasis studies.

Main Results:

• Identified several small molecules that bind to Ure2p.
• Discovered uretupamine, a compound that specifically activates the glucose-sensitive transcriptional pathway downstream of Ure2p.
• Confirmed uretupamine's specificity for Ure2p and its ability to modulate a specific subset of genes.

Conclusions:

• Diversity-oriented synthesis combined with small-molecule microarrays is an effective strategy for discovering protein-binding small molecules.
• Identified small molecules, like uretupamine, can precisely regulate specific protein functions and downstream biological pathways.