High-resolution chemical dissection of a model eukaryote reveals targets, pathways and gene functions
High-resolution chemical dissection of a model eukaryote reveals targets, pathways and gene functions
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DOI：
10.1016/j.micres.2013.11.004
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发表时间：
2014-01-01
影响因子：
6.7
通讯作者：
Movva, N. Rao
中科院分区：
文献类型：
--
作者：
Hoepfner, Dominic;Helliwell, Stephen B.;Movva, N. Rao
由于生物学的进化保守性，从真核模式生物的遗传研究中捕获的实验知识提供了对人类细胞途径和最终生理学的深入了解。酵母化学基因组学分析是一种用于注释细胞对小分子的反应的强有力的方法。使用优化的平台，我们提供了近1800个生物活性化合物的杂合和纯合缺失集合的相对灵敏度。数据质量使独特的见解的途径是敏感的和耐给定的扰动，如已知的和新的化合物证明。我们提出了抑制治疗相关的脂肪酸合成酶和去饱和酶（Fas1p和Ole1p）的新型化合物的例子，并展示了如何促进假设驱动的实验，以描绘化合物的作用机制。重要的是，测试化合物的规模和多样性产生了一个数据集，其中调节途径的数量接近饱和。该资源可用于绘制新的生物学连接，并确定未注释基因的功能。我们验证了由全局双向分层聚类生成的假设：（i）具有作用于微管或液泡ATP酶的类似作用机制的新型化合物，以及（ii）未注释的ORF YIL 060w，其在线粒体呼吸中起作用。最后，我们识别并表征了广泛使用的酵母缺失集合中的背景突变，这应该可以改善整个社区对过去和未来筛查的解释。这一全面的细胞反应资源使我们能够扩展对真核生物通路生物学的理解。(C)2013 Elsevier GmbH. All rights reserved.
Due to evolutionary conservation of biology, experimental knowledge captured from genetic studies in eukaryotic model organisms provides insight into human cellular pathways and ultimately physiology. Yeast chemogenomic profiling is a powerful approach for annotating cellular responses to small molecules. Using an optimized platform, we provide the relative sensitivities of the heterozygous and homozygous deletion collections for nearly 1800 biologically active compounds. The data quality enables unique insights into pathways that are sensitive and resistant to a given perturbation, as demonstrated with both known and novel compounds. We present examples of novel compounds that inhibit the therapeutically relevant fatty acid synthase and desaturase (Fas1p and Ole1p), and demonstrate how the individual profiles facilitate hypothesis-driven experiments to delineate compound mechanism of action. Importantly, the scale and diversity of tested compounds yields a dataset where the number of modulated pathways approaches saturation. This resource can be used to map novel biological connections, and also identify functions for unannotated genes. We validated hypotheses generated by global two-way hierarchical clustering of profiles for (i) novel compounds with a similar mechanism of action acting upon microtubules or vacuolar ATPases, and (ii) an un-annotated ORF, YIL060w, that plays a role in respiration in the mitochondria. Finally, we identify and characterize background mutations in the widely used yeast deletion collection which should improve the interpretation of past and future screens throughout the community. This comprehensive resource of cellular responses enables the expansion of our understanding of eukaryotic pathway biology. (C) 2013 Elsevier GmbH. All rights reserved.