Paper ID: 112
1 Institute for Biochemistry - ETH Zürich (Switzerland)
2 University of Fribourg (Switzerland)
Saccharomyces cerevisiae is a powerful model for systems genetics. We present a versatile, time- and labor-efficient method to functionally explore the yeast genome using saturated transposon mutagenesis coupled to high-throughput sequencing. SAturated Transposon Analysis in Yeast (SATAY) allows one-step mapping of all genetic loci in which transposons can insert without disrupting essential functions. SATAY is particularly suited to discover loci important for growth under various conditions.
SATAY (1) reveals positive and negative genetic interactions in single and multiple mutant strains, (2) can identify drug targets, (3) detects not only essential genes, but also essential protein domains, and (4) generates null and other informative alleles.
This method allowed us to identify genes acting in a pathway that uses vacuolar components to bypass ER-mitochondria contact sites. Moreover, in a screen for rapamycin-resistant mutants, we identified Pib2 as a regulator of TORC1. We show that Pib2 possesses two antagonistic TORC1-activating and -inhibiting activities located on opposite ends of the protein. Thus, SATAY allows to easily explore the yeast genome at unprecedented resolution and throughput.