Subcellular localization of the yeast proteome
Subcellular localization of the yeast proteome
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DOI：
10.1101/gad.970902
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发表时间：
2002-03-15
影响因子：
10.5
通讯作者：
Snyder, M
中科院分区：
文献类型：
--
作者：
Kumar, A;Agarwal, S;Snyder, M
蛋白质定位数据是有助于阐明真核蛋白功能的宝贵信息资源。在这里，我们报告了任何真核生物中蛋白质定位的第一个蛋白质组规模分析。使用定向的拓扑异构酶i介导的克隆策略和全基因组跨座子诱变，我们将表位标记为60％的酿酒酵母蛋白质组。通过标记基因产物的高通量免疫定位，我们确定了2744种酵母蛋白的亚细胞定位。通过使用贝叶斯形式主义的计算算法来推断这些数据，我们定义了酵母菌局的（所有6100个酵母蛋白的亚细胞分布）。我们将酵母蛋白质组估计为-5100可溶性蛋白和> 1000个跨膜蛋白。我们的结果表明，有47％的酵母蛋白是细胞质，13％线粒体，13％的外生型（包括内质网和分泌囊泡的蛋白质）和27％的核/核酸。通过使用减数分裂染色体的表面开发制剂，通过免疫定位进一步分析了核蛋白的子集。在这些蛋白质中，发现38％与染色体DNA相关。从核蛋白的表型分析确定的那样，对于孢子生存能力，34％是必不可少的 - 几个百分比几乎是整个蛋白质组观察到的两倍。总的来说，这项研究介绍了以前未知功能的955种蛋白质的实验得出的定位数据：酵母中所有功能未表征的蛋白质中的几乎一半。为了促进访问这些数据，我们在http://ygae.med.yale.edu上提供了一个可搜索的数据库，其中包含2900个荧光显微照片。
Protein localization data are a valuable information resource helpful in elucidating eukaryotic protein function. Here, we report the first proteome-scale analysis of protein localization within any eukaryote. Using directed topoisomerase I-mediated cloning strategies and genome-wide transposon mutagenesis, we have epitope-tagged 60% of the Saccharomyces cerevisiae proteome. By high-throughput immunolocalization of tagged gene products, we have determined the subcellular localization of 2744 yeast proteins. Extrapolating these data through a computational algorithm employing Bayesian formalism, we define the yeast localizome (the subcellular distribution of all 6100 yeast proteins). We estimate the yeast proteome to encompass -5100 soluble proteins and >1000 transmembrane proteins. Our results indicate that 47% of yeast proteins are cytoplasmic, 13% mitochondrial, 13% exocytic (including proteins of the endoplasmic reticulum and secretory vesicles), and 27% nuclear/nucleolar. A subset of nuclear proteins was further analyzed by immunolocalization using surface-spread preparations of meiotic chromosomes. Of these proteins, 38% were found associated with chromosomal DNA. As determined from phenotypic analyses of nuclear proteins, 34% are essential for spore viability-a percentage nearly twice as great as that observed for the proteome as a whole. In total, this study presents experimentally derived localization data for 955 proteins of previously unknown function: nearly half of all functionally uncharacterized proteins in yeast. To facilitate access to these data, we provide a searchable database featuring 2900 fluorescent micrographs at http://ygae.med.yale.edu.