Parallel chemical genetic and genome-wide RNAi screens identify cytokinesis inhibitors and targets
Parallel chemical genetic and genome-wide RNAi screens identify cytokinesis inhibitors and targets
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DOI：
10.1371/journal.pbio.0020379
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发表时间：
2004-12-01
期刊：
影响因子：
9.8
通讯作者：
Field, CM
中科院分区：
文献类型：
--
作者：
Eggert, US;Kiger, AA;Field, CM
细胞质分裂涉及细胞周期、细胞骨架和膜系统在时间和空间上的协调作用，以实现子细胞的分离。为了解剖细胞分裂机制，有一个完整的蛋白质目录和小分子工具来特异性地抑制它们，严格的时间控制是有用的。通过基于细胞的筛选寻找活性小分子需要确定其目标的困难步骤。我们在果蝇细胞中进行了平行的化学遗传和全基因组RNA干扰筛选，鉴定了50个细胞分裂小分子抑制剂和214个细胞分裂重要基因，包括Aurora B通路中的一个新蛋白（Borr）。通过比较小分子和RNAi表型，我们确定了一个抑制极光B激酶途径的小分子。我们的蛋白质列表为细胞分裂的系统解剖提供了一个起点，我们已经确定的各种小分子抑制剂将极大地促进这一方向。对Aurora B通路的解剖，我们发现了一个新的基因和一个特定的小分子抑制剂，应该会特别受益。我们的研究表明，平行RNA干扰和小分子筛选是识别活性小分子及其靶途径的一种普遍有用的方法。
Cytokinesis involves temporally and spatially coordinated action of the cell cycle and cytoskeletal and membrane systems to achieve separation of daughter cells. To dissect cytokinesis mechanisms it would be useful to have a complete catalog of the proteins involved, and small molecule tools for specifically inhibiting them with tight temporal control. Finding active small molecules by cell-based screening entails the difficult step of identifying their targets. We performed parallel chemical genetic and genome-wide RNA interference screens in Drosophila cells, identifying 50 small molecule inhibitors of cytokinesis and 214 genes important for cytokinesis, including a new protein in the Aurora B pathway (Borr). By comparing small molecule and RNAi phenotypes, we identified a small molecule that inhibits the Aurora B kinase pathway. Our protein list provides a starting point for systematic dissection of cytokinesis, a direction that will be greatly facilitated by also having diverse small molecule inhibitors, which we have identified. Dissection of the Aurora B pathway, where we found a new gene and a specific small molecule inhibitor, should benefit particularly. Our study shows that parallel RNA interference and small molecule screening is a generally useful approach to identifying active small molecules and their target pathways.