Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal 462023, India.
Department of Chemistry, Indian Institute of Science Education and Research, Bhopal 462023, India.
Eur J Pharmacol. 2014 Aug 5;736:77-85. doi: 10.1016/j.ejphar.2014.04.032. Epub 2014 May 2.
Ruthenium-based anti-cancer drugs have attracted increasing interest in the last 20 years. KP1019 is one of the ruthenium-containing compounds that has demonstrated anti-tumor activity against various cancers, and has been tested in several clinical trials. Despite its success, the mode of action of KP1019 is not well described. In the present study, we have used budding yeast Saccharomyces cerevisiae to elucidate the action of KP1019. We have found that KP1019 causes dose-dependent cell arrest in the S-phase of cell cycle. Furthermore, we have demonstrated for the first time that the yeast mitogen-activated protein (MAP) kinase Hog1 is essential for the cells in response to KP1019. Hog1 is rapidly phosphorylated upon treatment with KP1019, and the deletion of the HOG1 gene potentiates the growth inhibition effect of KP1019. Moreover, we also observed the up-regulation of glycerol-3-phosphate dehydrogenase 1 (GPD1) mRNA in response to KP1019 treatment, a factor that is essential for the hyperosmotic stress response. Our results also reveal that membrane-bound sensor proteins of high osmolarity glycerol (HOG) pathway are crucial for Hog1 phosphorylation in response to KP1019-induced stress. Furthermore, KP1019 has also been found to increase the accumulation of ubiquitinated proteins and deletion of several members of ubiquitination pathways conferred sensitivity for KP1019. The findings presented here strongly suggest the ability of KP1019 to activate Hog1 MAP kinase and induce protein ubiquitination, which may underlie the therapeutic potential of this compound. In summary, we have disclosed a novel mechanism of KP1019 activity.
钌基抗癌药物在过去 20 年中引起了越来越多的关注。KP1019 是一种含钌的化合物，它对各种癌症具有抗肿瘤活性，并在几项临床试验中进行了测试。尽管取得了成功，但 KP1019 的作用机制尚未得到很好的描述。在本研究中，我们使用 budding yeast Saccharomyces cerevisiae 来阐明 KP1019 的作用。我们发现 KP1019 导致细胞周期 S 期的细胞停滞呈剂量依赖性。此外，我们首次证明酵母有丝分裂原激活蛋白 (MAP) 激酶 Hog1 对细胞响应 KP1019 至关重要。KP1019 处理后 Hog1 迅速磷酸化，HOG1 基因缺失增强了 KP1019 的生长抑制作用。此外，我们还观察到甘油-3-磷酸脱氢酶 1 (GPD1) mRNA 的上调响应 KP1019 处理，这是高渗应激反应所必需的因素。我们的结果还表明，高渗透压甘油 (HOG) 途径的膜结合传感器蛋白对于 Hog1 磷酸化响应 KP1019 诱导的应激至关重要。此外，还发现 KP1019 增加了泛素化蛋白的积累，并且泛素化途径的几个成员的缺失赋予了对 KP1019 的敏感性。这里提出的发现强烈表明 KP1019 能够激活 Hog1 MAP 激酶并诱导蛋白质泛素化，这可能是该化合物治疗潜力的基础。总之，我们揭示了 KP1019 活性的一种新机制。
