Subcellular localization of the yeast proteome.
Anuj Kumar (1), Seema Agarwal (1), John Heyman (2), Amar Drawid
(3), Kei Cheung (4), Mark Gerstein (1), Shirleen Roeder (1), Michael
Snyder (1)
(1) MCD Biology, Yale University, 266 Whitney Avenue, New Haven, CT
06511, United States;
(2) Invitrogen Corporation, Carlsbad, CA 92008, USA;
(3) Dept. Mol. Biophys. & Biochem., Yale University, New Haven, CT
06520;
(4) Center for Medical Informatics, Dept. of Anesthesiology, Yale
University, School of Medicine, New Haven, CT 06510
Protein localization data is 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 this data
through a computational algorithm employing Bayesian formalism, we
define the yeast 'localizome' (the subcellular distribution of all 6100
yeast proteins). Our results indicate that 47% of yeast proteins are
cytoplasmic, 13% mitochondrial, 13% exocytic, and 27% nuclear. A subset
of nuclear proteins were further analyzed by immunolocalization using
surface-spread preparations of meiotic chromosomes. 38% of these
proteins were found associated with chromosomal DNA. As determined from
phenotypic analyses of nuclear proteins, 34% are essential for spore
viability -- a percentage nearly twice greater than 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 this data, we offer a searchable database
featuring 2900 fluorescent micrographs at ygac.med.yale.edu.
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