In 2014, the coal cleaning chemical 4-methylcyclohexane methanol (MCHM) spilled into the water supply for 300,000 West Virginians. Initial toxicology tests showed relatively mild results, but the underlying effects on cellular biology were underexplored. Treated wildtype yeast cells grew poorly, but there was only a small decrease in cell viability. Cell cycle analysis revealed an absence of cells in S phase within thirty minutes of treatment. Cells accumulated in G1 over a six-hour time course, indicating arrest instead of death. A genetic screen of the haploid knockout collection revealed 329 high confidence genes required for optimal growth in MCHM. These genes encode three major cell processes: mitochondrial gene expression/translation, the vacuolar ATPase, and aromatic amino acid biosynthesis. The transcriptome showed an upregulation of pleiotropic drug response genes and amino acid biosynthetic genes and downregulation in ribosome biosynthesis. Analysis of these datasets pointed to environmental stress response activation upon treatment. Overlap in datasets included the aromatic amino acid genes ARO1, ARO3, and four of the five TRP genes. This implicated nutrient deprivation as the signal for stress response. Excess supplementation of nutrients and amino acids did not improve growth on MCHM, so the source of nutrient deprivation signal is still unclear. Reactive oxygen species and DNA damage were directly detected with MCHM treatment, but timepoints showed these accumulated slower than cells arrested. We propose that wildtype cells arrest from nutrient deprivation and survive, accumulating oxidative damage through the implementation of robust environmental stress responses.

2014 年，煤炭清洁化学品 4-甲基环己烷甲醇 (MCHM) 泄漏到 30 万西弗吉尼亚州的供水系统中。最初的毒理学测试显示结果相对温和，但对细胞生物学的潜在影响尚未得到充分探索。处理后的野生型酵母细胞生长不良，但细胞活力仅略有下降。细胞周期分析显示在治疗后三十分钟内不存在处于S期的细胞。细胞在 6 小时的时间内积累在 G1 期，表明细胞停滞而不是死亡。单倍体敲除集合的遗传筛选揭示了 MCHM 最佳生长所需的 329 个高可信度基因。这些基因编码三个主要细胞过程：线粒体基因表达/翻译、液泡 ATP 酶和芳香氨基酸生物合成。转录组显示多效药物反应基因和氨基酸生物合成基因上调，核糖体生物合成下调。对这些数据集的分析表明治疗后环境应激反应被激活。数据集中的重叠包括芳香族氨基酸基因ARO1 、ARO3 和五个 TRP 基因中的四个。这表明营养缺乏是应激反应的信号。过量补充营养物质和氨基酸并不能改善MCHM的生长，因此营养缺乏信号的来源仍不清楚。 MCHM 处理可直接检测到活性氧和 DNA 损伤，但时间点显示这些积累速度比细胞停滞的速度慢。我们认为野生型细胞会因营养缺乏而停滞并存活下来，通过实施强有力的环境应激反应来积累氧化损伤。