In the aftermath of the January 2014 MCHM spill into the Elk River of Kanawha county, West Virginia, the lack of knowledge concerning the mode of action of this chemical within the cell became clear. The budding yeast is a key model organism with conserved eukaryotic biochemical and genetic pathways, as well as convenient functional genomics tools with which to identify how the cell responds to chemicals. Investigations into MCHM cellular response included growth assays of wild type strains, viability assays, growth assays of knockout strains, RNA-seq analysis, quantitative trait loci, and GFP-labeled protein localization microscopy. Viability and growth assays of wild type yeast revealed sensitivity to MCHM in the concentration range of approximately 0.04-0.06%, though defect in growth appears to be from inhibition of growth instead of lethality. RNA-seq analysis for differential expression of genes with MCHM treatment indicated that genes for transcription factors and ABC transporters in the pleiotropic drug response are affected. Knockout strains for drug response and oxidative stress genes were also grown in MCHM, revealing several mutants in these pathways were sensitive, and therefore required for MCHM response. The RNA-seq data also pointed to the inositol biosynthetic pathway, which is normally tightly regulated by a certain set of proteins. One gene in particular, the major negative regulator of UAS INO genes, opi1, showed sensitivity to MCHM. Many of the genes regulated by the Opi1 protein are downregulated by MCHM. Microscopy of the GFP-labeled Opi1p revealed relocalization of the negative inhibitor protein into the nucleus due to MCHM treatment in media lacking inositol, the normal signal for relocalization. Future investigation into the effects of MCHM must involve investigation of pleiotropic drug response, oxidative stress, and the inositol biosynthetic pathway to reveal mechanisms of toxicity and targets of treatment for exposure.