High-resolution chemical dissection of a model eukaryote reveals targets, pathways and gene functions

Microbiol Res. 2014 Feb-Mar;169(2-3):107-20. doi: 10.1016/j.micres.2013.11.004. Epub 2013 Dec 1.

Abstract

Due to evolutionary conservation of biology, experimental knowledge captured from genetic studies in eukaryotic model organisms provides insight into human cellular pathways and ultimately physiology. Yeast chemogenomic profiling is a powerful approach for annotating cellular responses to small molecules. Using an optimized platform, we provide the relative sensitivities of the heterozygous and homozygous deletion collections for nearly 1800 biologically active compounds. The data quality enables unique insights into pathways that are sensitive and resistant to a given perturbation, as demonstrated with both known and novel compounds. We present examples of novel compounds that inhibit the therapeutically relevant fatty acid synthase and desaturase (Fas1p and Ole1p), and demonstrate how the individual profiles facilitate hypothesis-driven experiments to delineate compound mechanism of action. Importantly, the scale and diversity of tested compounds yields a dataset where the number of modulated pathways approaches saturation. This resource can be used to map novel biological connections, and also identify functions for unannotated genes. We validated hypotheses generated by global two-way hierarchical clustering of profiles for (i) novel compounds with a similar mechanism of action acting upon microtubules or vacuolar ATPases, and (ii) an un-annotated ORF, YIL060w, that plays a role in respiration in the mitochondria. Finally, we identify and characterize background mutations in the widely used yeast deletion collection which should improve the interpretation of past and future screens throughout the community. This comprehensive resource of cellular responses enables the expansion of our understanding of eukaryotic pathway biology.

Keywords: Chemogenomic profiling; Cmpd; FAS; HIP; HIP HOP profiling; HOP; Mechanism of action; MoA; Target identification; Yeast; compound; fatty acid synthase; haploinsufficiency profiling; homozygous profiling; mechanism of action.

MeSH terms

  • Antifungal Agents / pharmacology
  • Biosynthetic Pathways
  • Drug Resistance, Fungal
  • Gene Expression Regulation, Fungal
  • High-Throughput Screening Assays
  • Molecular Sequence Data
  • Phylogeny
  • Saccharomyces cerevisiae / chemistry*
  • Saccharomyces cerevisiae / classification
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism

Substances

  • Antifungal Agents
  • Saccharomyces cerevisiae Proteins