Functional mapping of yeast genomes by saturated transposition

Agnès H Michel, Riko Hatakeyama, Philipp Kimmig, Meret Arter, Matthias Peter, Joao Matos, Claudio De Virgilio, Benoît Kornmann* (Corresponding Author)

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

49 Citations (Scopus)

Abstract

Yeast is a powerful model for systems genetics. We present a versatile, time- and labor-efficient method to functionally explore the Saccharomyces cerevisiae 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.

Original languageEnglish
Article numbere23570
Number of pages28
JournaleLife
Volume6
Early online date8 May 2017
DOIs
Publication statusPublished - 9 Jun 2017

Keywords

  • DNA Transposable Elements
  • Genes, Fungal
  • Genetics, Microbial/methods
  • Genome, Fungal
  • Molecular Sequence Annotation/methods
  • Mutagenesis, Insertional/methods
  • Saccharomyces cerevisiae/genetics
  • Sequence Analysis, DNA

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