S. cerevisiae exists as a haploid or as a diploid. Deleting 1 of the 2 copies of a gene in diploid strains can reduce its expression, and a set

of ~6,000 heterozygous diploid strains covering nearly all essential and nonessential genes is available. Complete deletion of nonessential genes eliminates their expression and sets of ~4,900 haploid and homozygous diploid deletion mutants are also available. S. cerevisiae can be easily transformed and increased gene expression can be achieved by introducing plasmids containing genomic DNA fragments or gene-coding regions controlled by inducible promoters [3]. The unicellular nature of yeast and its ability to grow on liquid or solid media also make it amenable to high-throughput drug studies. A number Y-27632 manufacturer of studies have shown that reducing the copy number of essential or nonessential

genes from 2 to 1 in diploid cells may increase the sensitivity of the cell to a drug (termed drug-induced haploinsufficiency) and can point to candidate target genes [4–6]. Haploid or homozygous diploid deletion collections contain only deletions of nonessential genes. Screening these collections for hypersensitivity to a small molecule reveals genes that buffer the drug target pathway, not the direct drug targets and comparison of the profile of chemical-genetic synthetic lethality with a compendium of chemical-genetic or genetic interaction profiles can aid in deciphering its targets [7, 8]. Increased gene expression can lead to suppression of drug sensitivity and also reveal learn more target genes [3, 9]. Studies of the mechanism of action of drugs using genome-wide approaches in yeast have tended to focus on 1 of these 3 approaches [3, 5, 8]. While each generally reveals important clues, they draw only a partial picture of the mechanism of action of chemicals. For example, a drug-induced haploinsufficiency screen of the cancer cell invasion inhibitor dihydromotuporamine stiripentol C (dhMotC) showed that the compound targets sphingolipid biosynthesis and affects the actin cytoskeleton

[6], but did not reveal whether other cellular functions were affected and gave no indication of cell death mechanisms involved. Genome-wide studies of drug mechanism of action have mainly concentrated on nuclear-encoded genes. Genes encoded by mitochondrial DNA, which include components of the mitochondrial translational machinery and 8 mitochondrial proteins, have not received as much attention. Yet mitochondria are recognized as important regulators of cell death in addition to their central role in energy production [10]. Although yeast displays only some of the characteristics of apoptosis described in humans, many cellular features of the cell death pathway in mammalian cells have been 17DMAG concentration identified in yeast [11].