Genome-wide SWAp-tag yeast libraries for proteome exploration

Yeast libraries revolutionized the systematic study of cell biology. To extensively increase the number of such libraries and the type of information that can be gleaned from them, we previously devised the SWAp-Tag (SWAT) approach that enables rapid, easy and efficient creation of yeast strain collections. Here we present the construction and investigation of a full genome library of ~5500 strains carrying the SWAT NOP1promoter-GFP module at the N terminus of proteins, as well as its use in creating six additional libraries that either restore the native regulation, create an overexpression library with a Cherry tag or enable protein complementation assays from two fragments of an enzyme or fluorophore. We show methods to utilize these SWAT collections to systematically characterize the yeast proteome on multiple levels spanning protein abundance, localization, topology and interactions. Our findings demonstrate how diverse full-genome SWAT libraries facilitate obtaining insights into numerous aspects of the proteome.