title: Subcellular localization of the yeast proteome.
abstract: 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.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520, USA.
authors.forename: Anuj
authors.initials: A
authors.lastname: Kumar
authors.forename: Seema
authors.initials: S
authors.lastname: Agarwal
authors.forename: John A
authors.initials: JA
authors.lastname: Heyman
authors.forename: Sandra
authors.initials: S
authors.lastname: Matson
authors.forename: Matthew
authors.initials: M
authors.lastname: Heidtman
authors.forename: Stacy
authors.initials: S
authors.lastname: Piccirillo
authors.forename: Lara
authors.initials: L
authors.lastname: Umansky
authors.forename: Amar
authors.initials: A
authors.lastname: Drawid
authors.forename: Ronald
authors.initials: R
authors.lastname: Jansen
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Kei-Hoi
authors.initials: KH
authors.lastname: Cheung
authors.forename: Perry
authors.initials: P
authors.lastname: Miller
authors.forename: Mark
authors.initials: M
authors.lastname: Gerstein
authors.forename: G Shirleen
authors.initials: GS
authors.lastname: Roeder
authors.forename: Michael
authors.initials: M
authors.lastname: Snyder
mesh_terms: D000465:Algorithms; D002467:Cell Nucleus / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D019992:Databases as Topic; D000939:Epitopes; D016681:Genome, Fungal; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D008957:Models, Genetic; D016296:Mutagenesis; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012984:Software
chemical_list: D000939:Epitopes