Functional mapping of yeast genomes by saturated transposition.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 28481201.
- Also identified by DOI 10.7554/eLife.23570 and PMC identifier 5466422.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Yeast is a powerful model for systems genetics. We present a versatile, time- and labor-efficient method to functionally explore the <i>Saccharomyces cerevisiae</i> 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, (4) generates both null and other informative alleles. In a SATAY screen for rapamycin-resistant mutants, we identify Pib2 (PhosphoInositide-Binding 2) as a master regulator of TORC1. We describe two antagonistic TORC1-activating and -inhibiting activities located on opposite ends of Pib2. Thus, SATAY allows to easily explore the yeast genome at unprecedented resolution and throughput.
Medical subject headings
- Genes, Fungal
- Genetics, Microbial
- Molecular Sequence Annotation
- Mutagenesis, Insertional
- Saccharomyces cerevisiae