Exploration of essential gene functions via titratable promoter alleles
Exploration of essential gene functions via titratable promoter alleles
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DOI：
10.1016/j.cell.2004.06.013
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发表时间：
2004-07-09
期刊：
影响因子：
64.5
通讯作者：
Hughes, TR
中科院分区：
文献类型：
--
作者：
Mnaimneh, S;Davierwala, AP;Hughes, TR
近20%的酵母基因是生存所必需的，这阻碍了基因敲除的遗传分析。我们为超过三分之二的必需酵母基因创建了启动子关闭菌株，并对其进行形态分析，大小分析，药物敏感性筛选和微阵列表达谱分析。然后，我们使用这些数据纲要来询问哪些表型特征表征了不同的功能类别，并使用这些特征来推断未表征基因的潜在功能。我们鉴定了参与核糖体生物发生（HAS 1、URB 1和URB 2）、蛋白质分泌（SEC 39）、线粒体输入（MIM 1）和tRNA充电（GSN 1）的基因。此外，观察到核糖体生物合成和蛋白酶体的明显负反馈转录调节。我们还表明，这些菌株是兼容的自动遗传分析。这项研究强调了分析突变表型的重要性，并提供了一个资源，以补充酵母敲除收集。
Nearly 20% of yeast genes are required for viability, hindering genetic analysis with knockouts. We created promoter-shutoff strains for over two-thirds of all essential yeast genes and subjected them to morphological analysis, size profiling, drug sensitivity screening, and microarray expression profiling. We then used this compendium of data to ask which phenotypic features characterized different functional classes and used these to infer potential functions for uncharacterized genes. We identified genes involved in ribosome biogenesis (HAS1, URB1, and URB2), protein secretion (SEC39), mitochondrial import (MIM1), and tRNA charging (GSN1). In addition, apparent negative feedback transcriptional regulation of both ribosome biogenesis and the proteasome was observed. We furthermore show that these strains are compatible with automated genetic analysis. This study underscores the importance of analyzing mutant phenotypes and provides a resource to complement the yeast knockout collection.