AIMMS, Division of Molecular Toxicology, Department of Chemistry and Pharmaceutical Sciences, VU University Amsterdam, De Boelelaan 1083, 1081 HZ, Amsterdam, The Netherlands.
AIMMS, Department of Molecular Cell Biology, Section Genetics, VU University Amsterdam, De Boelelaan 1083, 1081 HZ, Amsterdam, The Netherlands.
Amino Acids. 2018 Oct;50(10):1377-1390. doi: 10.1007/s00726-018-2613-8. Epub 2018 Jul 5.
In yeast, toxicity of acetaminophen (APAP), a frequently used analgesic and antipyretic drug, depends on ubiquitin-controlled processes. Previously, we showed a remarkable overlap in toxicity profiles between APAP and tyrosine, and a similarity with drugs like rapamycin and quinine, which induce degradation of the amino acid permease Tat2. Therefore, we investigated in yeast whether APAP reduced the expression levels of amino acid permeases. The protein levels of Tat2, Tat1, Mup1 and Hip1 were reduced, while the expression of the general permease Gap1 was increased, consistent with a nutrient starvation response. Overexpression of Tat1 and Tat2, but not Mup1, Hip1 and Gap1 conferred resistance to APAP. A tryptophan auxotrophic strain trp1Δ was more sensitive to APAP than wild-type and addition of tryptophan completely restored the growth restriction of trp1∆ upon APAP exposure, while tyrosine had an additive effect on APAP toxicity. Furthermore, intracellular aromatic amino acid concentrations were reduced upon APAP exposure. This effect was less prominent in ubiquitin-deficient yeast strains that were APAP resistant and showed a reduced degradation of high affinity amino acid permeases. APAP-induced changes in intracellular amino acid concentrations were also detected in hepatoma HepG2 cells indicating significance for humans.
在酵母中,常用的解热镇痛药对乙酰氨基酚(APAP)的毒性取决于泛素控制的过程。之前,我们发现 APAP 的毒性谱与酪氨酸有显著重叠,与雷帕霉素和奎宁等药物的相似性,这些药物诱导氨基酸转运蛋白 Tat2 的降解。因此,我们在酵母中研究了 APAP 是否降低了氨基酸转运蛋白的表达水平。Tat2、Tat1、Mup1 和 Hip1 的蛋白水平降低,而一般通透性 Gap1 的表达增加,与营养饥饿反应一致。Tat1 和 Tat2 的过表达,但不是 Mup1、Hip1 和 Gap1,赋予了对 APAP 的抗性。色氨酸营养缺陷型菌株 trp1Δ 比野生型对 APAP 更敏感,添加色氨酸完全恢复了 trp1Δ 在 APAP 暴露下的生长限制,而酪氨酸对 APAP 毒性有相加作用。此外,APAP 暴露后细胞内芳香族氨基酸浓度降低。在对 APAP 有抗性且高亲和力氨基酸转运蛋白降解减少的泛素缺陷酵母菌株中,这种影响不太明显。在肝癌 HepG2 细胞中也检测到了 APAP 诱导的细胞内氨基酸浓度变化,表明对人类有重要意义。