Abstract
Protein complexes are key molecular entities that integrate multiple gene products to perform cellular functions. Here we report the first genome-wide screen for complexes in an organism, budding yeast, using affinity purification and mass spectrometry. Through systematic tagging of open reading frames (ORFs), the majority of complexes were purified several times, suggesting screen saturation. The richness of the data set enabled a de novo characterization of the composition and organization of the cellular machinery. The ensemble of cellular proteins partitions into 491 complexes, of which 257 are novel, that differentially combine with additional attachment proteins or protein modules to enable a diversification of potential functions. Support for this modular organization of the proteome comes from integration with available data on expression, localization, function, evolutionary conservation, protein structure and binary interactions. This study provides the largest collection of physically determined eukaryotic cellular machines so far and a platform for biological data integration and modelling.
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Acknowledgements
We thank C. Cohen, S. Artavanis-Tsakonas, B. Seraphin and L. Serrano for support and suggestions throughout the work, and F. Weisbrodt for assistance with the graphics.
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Purification and complex data have been deposited at the IntAct database (http://www.ebi.ac.uk/intact/) with accession numbers EBI-768904 (purifications) and EBI-765905 (author inferred complexes). The data, including the MS protein identifications, are accessible at http://yeast-complexes.embl.de, and the yeast strains are available from Euroscarf (http://web.uni-frankfurt.de/fb15/mikro/euroscarf/col_index.html). Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
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Supplementary Notes
This file contains Supplementary Data and Supplementary Methods, including a detailed description of the screen for protein complexes with Supplementary Data on the proteome coverage. There is also a detailed description of the biochemical, mass spectrometry and bioinformatics methods. This file also contains Supplementary Figures 1–9 as supports to the points described above. (PDF 1196 kb)
Supplementary Table 1
List of all purifications and proteins retrieved. (PDF 344 kb)
Supplementary Table 2
List of all protein complexes. (PDF 202 kb)
Supplementary Table 3
List of all protein modules. (PDF 64 kb)
Supplementary Table 4
List of protein complexes purified only once and producing a signal too weak to be automatically deduced. (PDF 62 kb)
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Gavin, AC., Aloy, P., Grandi, P. et al. Proteome survey reveals modularity of the yeast cell machinery. Nature 440, 631–636 (2006). https://doi.org/10.1038/nature04532
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DOI: https://doi.org/10.1038/nature04532
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