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Subcellular localization of the yeast proteome.

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 approximately 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://ygac.med.yale.edu.

Pubmed ID: 11914276

Authors

  • Kumar A
  • Agarwal S
  • Heyman JA
  • Matson S
  • Heidtman M
  • Piccirillo S
  • Umansky L
  • Drawid A
  • Jansen R
  • Liu Y
  • Cheung KH
  • Miller P
  • Gerstein M
  • Roeder GS
  • Snyder M

Journal

Genes & development

Publication Data

March 15, 2002

Associated Grants

  • Agency: NCI NIH HHS, Id: R01-CA77808

Mesh Terms

  • Algorithms
  • Cell Nucleus
  • Chromosomes
  • Cytoplasm
  • Databases as Topic
  • Epitopes
  • Genome, Fungal
  • Microscopy, Fluorescence
  • Mitochondria
  • Models, Genetic
  • Mutagenesis
  • Phenotype
  • Saccharomyces cerevisiae
  • Software