Assigning Function to Yeast Proteins by Integration of Technologies
Tony R. Hazbun1,2, Lars Malmström3, Scott Anderson4, Beth J. Graczyk3, Bethany Fox3, Michael Riffle3, Bryan A. Sundin3, J. Derringer Aranda2, W. Hayes McDonald4, Chui-hwei Chun3, Brian E. Snydsman3, Phillip Bradley3, Eric G. D. Muller3, Stanley Fields1,2, David Baker1,3, John R. Yates III4 and Trisha N. Davis3*
1Howard Hughes Medical Institute, 2Departments of Genome Sciences and Medicine, 3Department of Biochemistry, University of Washington, Seattle, WA, 4Department of Cell Biology, Scripps Research Institute, La Jolla, CA
Interpreting genome sequences requires the functional analysis of thousands of predicted proteins, many of which are uncharacterized and without obvious homologues. 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 co-purifying 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 classify genes.
How To Use This Site:
This site provides the ability to search for all of the data we found for a particular ORF or gene. To search, simply type in the name of the ORF or gene in the search box near the top of every page. This search is not limited to the 100 essential uncharacterized ORFs we studied. You are able to view and download all of our mass spectrometry, fluorescence microscopy, yeast two-hybrid and protein structure prediction data for any ORF for which we have experimental data.
In addition, we provide three tables to summarize our findings for the 100 essential uncharacterized ORFs.
Finally, the About YRC page will give you a brief description of the Yeast Resource Center, and provide a link to our official web site.