Imipridone Anticancer Compounds Ectopically Activate the ClpP Protease and Represent a New Scaffold for Antibiotic Development
Imipridone Anticancer Compounds Ectopically Activate the ClpP Protease and Represent a New Scaffold for Antibiotic Development
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DOI：
10.1534/genetics.119.302851
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发表时间：
2020-04-01
期刊：
影响因子：
3.3
通讯作者：
Tyers, Mike
中科院分区：
文献类型：
--
作者：
Jacques, Samuel;van der Sloot, Almer M.;Tyers, Mike
系统的遗传相互作用谱可以揭示生物活性化合物的作用机制。目前正在进行癌症临床试验的依米立酮ONC 201被认为具有多种不同的靶点。为了研究依米立酮作用的遗传依赖性，我们在ONC 201或其更有效的类似物ONC 212存在下筛选了全基因组成簇的规则间隔短回文重复序列（CRISPR）敲除文库。线粒体基质蛋白酶CLPP或线粒体中间体肽酶MIPEP的损失赋予了对这两种化合物的强抗性。生物化学和替代遗传分析表明，impridone直接激活CLPP和MIPEP是必要的蛋白水解成熟的CLPP成催化活性的形式。用ONC 212处理的细胞的定量蛋白质组学分析揭示了许多线粒体以及非线粒体蛋白质的降解。由于ClpP从细菌到人类的保守性，我们发现在生物化学和遗传测定中，依米普利酮也激活来自大肠杆菌、枯草芽孢杆菌和金黄色葡萄球菌的ClpP。ONC 212和已知的细菌ClpP激活剂酰基缩酚酸肽-4（ADEP 4）在S.金黄色。ONC 212抑制了许多革兰氏阳性（S. aureus，B. B.枯草杆菌和屎肠球菌）和革兰氏阴性菌（E.大肠杆菌和淋病奈瑟菌）。此外，ONC 212增强了利福平根除耐药沙门氏菌的能力。金黄色葡萄球菌持留细胞。这些结果揭示了在人类细胞中的依米普利酮作用的遗传依赖性，并将依米普利酮支架确定为抗生素开发的新切入点。依米普利酮ONC 201和ONC 212选择性地杀死癌细胞，并被认为具有多种作用机制。全基因组CRISPR敲除筛选显示，线粒体蛋白酶CLPP和MIPEP的缺失赋予了对这两种化合物的强烈抗性。
Systematic genetic interaction profiles can reveal the mechanisms-of-action of bioactive compounds. The imipridone ONC201, which is currently in cancer clinical trials, has been ascribed a variety of different targets. To investigate the genetic dependencies of imipridone action, we screened a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) knockout library in the presence of either ONC201 or its more potent analog ONC212. Loss of the mitochondrial matrix protease CLPP or the mitochondrial intermediate peptidase MIPEP conferred strong resistance to both compounds. Biochemical and surrogate genetic assays showed that impridones directly activate CLPP and that MIPEP is necessary for proteolytic maturation of CLPP into a catalytically competent form. Quantitative proteomic analysis of cells treated with ONC212 revealed degradation of many mitochondrial as well as nonmitochondrial proteins. Prompted by the conservation of ClpP from bacteria to humans, we found that the imipridones also activate ClpP from Escherichia coli, Bacillus subtilis, and Staphylococcus aureus in biochemical and genetic assays. ONC212 and acyldepsipeptide-4 (ADEP4), a known activator of bacterial ClpP, caused similar proteome-wide degradation profiles in S. aureus. ONC212 suppressed the proliferation of a number of Gram-positive (S. aureus, B. subtilis, and Enterococcus faecium) and Gram-negative species (E. coli and Neisseria gonorrhoeae). Moreover, ONC212 enhanced the ability of rifampin to eradicate antibiotic-tolerant S. aureus persister cells. These results reveal the genetic dependencies of imipridone action in human cells and identify the imipridone scaffold as a new entry point for antibiotic development.The imipridones ONC201 and ONC212 selectively kill cancer cells and have been ascribed multiple mechanisms-of-action. Genome-wide CRISPR knockout screens revealed that loss of the mitochondrial proteases CLPP and MIPEP confer strong resistance to both compounds...