Genome-wide bimolecular fluorescence complementation analysis of SUMO interactome in yeast

Min-Kyung Sung; Gyubum Lim; Dae-Gwan Yi; Yeon Ji Chang; Eun Bin Yang; KiYoung Lee; Won-Ki Huh

doi:10.1101/gr.148346.112

Genome-wide bimolecular fluorescence complementation analysis of SUMO interactome in yeast

¹Department of Biological Sciences, Seoul National University, Seoul 151-747, Republic of Korea;
²Research Center for Functional Cellulomics and Institute of Microbiology, Seoul National University, Seoul 151-742, Republic of Korea;
³Department of Biomedical Informatics, Ajou University School of Medicine, Suwon 443-749, Republic of Korea

Abstract

The definition of protein–protein interactions (PPIs) in the natural cellular context is essential for properly understanding various biological processes. So far, however, most large-scale PPI analyses have not been performed in the natural cellular context. Here, we describe the construction of a Saccharomyces cerevisiae fusion library in which each endogenous gene is C-terminally tagged with the N-terminal fragment of Venus (VN) for a genome-wide bimolecular fluorescence complementation assay, a powerful technique for identifying PPIs in living cells. We illustrate the utility of the VN fusion library by systematically analyzing the interactome of the small ubiquitin-related modifier (SUMO) and provide previously unavailable information on the subcellular localization, types, and protease dependence of SUMO interactions. Our data set is highly complementary to the existing data sets and represents a useful resource for expanding the understanding of the physiological roles of SUMO. In addition, the VN fusion library provides a useful research tool that makes it feasible to systematically analyze PPIs in the natural cellular context.

Footnotes

↵4 Corresponding author

E-mail wkh{at}snu.ac.kr
[Supplemental material is available for this article.]
Article published online before print. Article, supplemental material, and publication date are at http://www.genome.org/cgi/doi/10.1101/gr.148346.112.

Received August 22, 2012.
Accepted February 7, 2013.

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