title: Assigning function to yeast proteins by integration of technologies.
abstract: Interpreting genome sequences requires the functional analysis of thousands of predicted proteins, many of which are uncharacterized and without obvious homologs. To assess whether the roles of large sets of uncharacterized genes can be assigned by targeted application of a suite of technologies, we used four complementary protein-based methods to analyze a set of 100 uncharacterized but essential open reading frames (ORFs) of the yeast Saccharomyces cerevisiae. These proteins were subjected to affinity purification and mass spectrometry analysis to identify copurifying proteins, two-hybrid analysis to identify interacting proteins, fluorescence microscopy to localize the proteins, and structure prediction methodology to predict structural domains or identify remote homologies. Integration of the data assigned function to 48 ORFs using at least two of the Gene Ontology (GO) categories of biological process, molecular function, and cellular component; 77 ORFs were annotated by at least one method. This combination of technologies, coupled with annotation using GO, is a powerful approach to classifying genes.
authors.affiliation: Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
authors.forename: Tony R
authors.initials: TR
authors.lastname: Hazbun
authors.forename: Lars
authors.initials: L
authors.lastname: Malmström
authors.forename: Scott
authors.initials: S
authors.lastname: Anderson
authors.forename: Beth J
authors.initials: BJ
authors.lastname: Graczyk
authors.forename: Bethany
authors.initials: B
authors.lastname: Fox
authors.forename: Michael
authors.initials: M
authors.lastname: Riffle
authors.forename: Bryan A
authors.initials: BA
authors.lastname: Sundin
authors.forename: J Derringer
authors.initials: JD
authors.lastname: Aranda
authors.forename: W Hayes
authors.initials: WH
authors.lastname: McDonald
authors.forename: Chun-Hwei
authors.initials: CH
authors.lastname: Chiu
authors.forename: Brian E
authors.initials: BE
authors.lastname: Snydsman
authors.forename: Phillip
authors.initials: P
authors.lastname: Bradley
authors.forename: Eric G D
authors.initials: EG
authors.lastname: Muller
authors.forename: Stanley
authors.initials: S
authors.lastname: Fields
authors.forename: David
authors.initials: D
authors.lastname: Baker
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.forename: Trisha N
authors.initials: TN
authors.lastname: Davis
mesh_terms: D019295:Computational Biology; D016681:Genome, Fungal; D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D020543:Proteome / Q000032:analysis; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins