RT Journal
A1 Suter, Bernhard
A1 Fetchko, Michael J.
A1 Imhof, Ralph
A1 Graham, Christopher I.
A1 Stoffel-Studer, Ingrid
A1 Zbinden, Caroline
A1 Raghavan, Maanasa
A1 Lopez, Lianet
A1 Beneti, Lucija
A1 Hort, Jacqueline
A1 Fillingham, Jeffrey
A1 Greenblatt, Jack F.
A1 Giaever, Guri
A1 Nislow, Corey
A1 Stagljar, Igor
T1 Examining protein–protein interactions using endogenously tagged yeast arrays: The Cross-and-Capture system
JF Genome Research 
JO Genome Research 
YR 2007 
FD December 01 
VO 17 
IS 12 
SP 000 
OP 000 
DO 10.1101/gr.6667007 
UL http://genome.cshlp.org/content/early/2007/11/07/gr.6667007.abstract 
AB Comprehensive approaches to detect protein–protein interactions (PPIs) have been most successful in the yeast model system. Here we present “Cross-and-Capture,” a novel assay for rapid, sensitive assessment of PPIs via pulldown of differently tagged yeast strain arrays. About 500 yeast genes that function in DNA replication, repair, and recombination and nuclear proteins of unknown function were chromosomally tagged with six histidine residues or triple VSV epitopes. We demonstrate that the assay can interrogate a wide range of previously known protein complexes with increased resolution and sensitivity. Furthermore, we use “Cross-and-Capture” to identify two novel protein complexes: Rtt101p–Mms1p and Sae2p–Mre11p. The Rtt101p–Mms1p interaction was subsequently characterized by genetic and functional analyses. Our studies establish the “Cross-and-Capture” assay as a novel, versatile tool that provides a valuable complement for the next generation of yeast proteomic studies.