title: Genome-wide bimolecular fluorescence complementation analysis of SUMO interactome in yeast.
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.
authors: Department of Biological Sciences, Seoul National University, Seoul 151-747, Republic of Korea.
authors: Min-Kyung
authors: MK
authors: Sung
authors: Gyubum
authors: G
authors: Lim
authors: Dae-Gwan
authors: DG
authors: Yi
authors: Yeon Ji
authors: YJ
authors: Chang
authors: Eun Bin
authors: EB
authors: Yang
authors: Kiyoung
authors: K
authors: Lee
authors: Won-Ki
authors: WK
authors: Huh
mesh_terms: D002453:Cell Cycle / Q000235:genetics; D019295:Computational Biology / Q000379:methods; D015723:Gene Library; D023061:Gene Order; D005816:Genetic Complementation Test; D055106:Genome-Wide Association Study / Q000379:methods; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D015203:Reproducibility of Results; D025842:SUMO-1 Protein / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism*
chemical_list: D025842:SUMO-1 Protein; D029701:Saccharomyces cerevisiae Proteins
references: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10340-4
references: 7937952
references: Science. 2008 Jun 13;320(5882):1465-70
references: 18467557
references: Mol Cell Biol. 2000 Apr;20(7):2367-77
references: 10713161
references: Proc Natl Acad Sci U S A. 2004 Oct 5;101(40):14373-8
references: 15388847
references: Nature. 2003 Oct 16;425(6959):737-41
references: 14562106
references: Annu Rev Cell Dev Biol. 2006;22:159-80
references: 16753028
references: Nature. 2000 Feb 10;403(6770):623-7
references: 10688190
references: J Biol Chem. 2004 Jul 30;279(31):32262-8
references: 15166219
references: Nature. 2002 Jan 10;415(6868):180-3
references: 11805837
references: Methods Enzymol. 2002;350:3-41
references: 12073320
references: Nature. 1989 Jul 20;340(6230):245-6
references: 2547163
references: J Biol Chem. 2004 Oct 1;279(40):41346-51
references: 15292183
references: Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3469-74
references: 11904411
references: Proc Natl Acad Sci U S A. 1999 May 11;96(10):5394-9
references: 10318894
references: Nature. 1999 Mar 18;398(6724):246-51
references: 10094048
references: Proc Natl Acad Sci U S A. 2004 Oct 12;101(41):14782-7
references: 15469925
references: Annu Rev Biophys. 2008;37:465-87
references: 18573091
references: Eukaryot Cell. 2007 Mar;6(3):378-87
references: 17220465
references: Anal Chem. 2003 Apr 1;75(7):1584-9
references: 12705589
references: Nature. 2003 Oct 16;425(6959):686-91
references: 14562095
references: Mol Cell Proteomics. 2005 Mar;4(3):246-54
references: 15542864
references: Nat Rev Mol Cell Biol. 2007 Dec;8(12):947-56
references: 18000527
references: Methods Cell Biol. 1999;58:293-314
references: 9891388
references: Trends Cell Biol. 1999 Feb;9(2):57-60
references: 10087619
references: Nature. 2006 Mar 30;440(7084):637-43
references: 16554755
references: EMBO J. 2005 Apr 6;24(7):1440-52
references: 15775961
references: Nat Biotechnol. 2003 May;21(5):539-45
references: 12692560
references: Nature. 1995 Jan 5;373(6509):78-81
references: 7800043
references: Nature. 2002 Jan 10;415(6868):141-7
references: 11805826
references: Nature. 2006 Mar 30;440(7084):631-6
references: 16429126
references: J Biol Chem. 2005 Feb 11;280(6):4102-10
references: 15590687
references: Yeast. 2008 Apr;25(4):301-11
references: 18350525
references: Mol Cell. 2009 Jan 16;33(1):124-35
references: 19150434
references: Yeast. 2007 Sep;24(9):767-75
references: 17534848
references: J Microbiol Methods. 2010 Nov;83(2):194-201
references: 20828586
references: J Biol Chem. 2005 Jan 14;280(2):1438-47
references: 15509571
references: J Biol Chem. 2004 Oct 29;279(44):45662-8
references: 15326169
references: Mol Cell Proteomics. 2005 Jan;4(1):73-83
references: 15596868
references: Nat Cell Biol. 2007 Oct;9(10):1184-91
references: 17828247
references: Mol Cell. 2002 Apr;9(4):789-98
references: 11983170
references: Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4569-74
references: 11283351
references: Mol Cell. 2003 Nov;12(5):1287-300
references: 14636585
references: Nat Biotechnol. 1999 Oct;17(10):1030-2
references: 10504710