Functional mapping of yeast genomes by saturated transposition
- ETH Zurich, Switzerland
- University of Fribourg, Switzerland
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.
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Funding
European Commission (337906-OrgaNet)
- Benoît Kornmann
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (PP00P3_13365)
- Benoît Kornmann
Human Frontier Science Program
- Philipp Kimmig
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (310030_166474)
- Claudio De Virgilio
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (31003A_153058)
- Joao Matos
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (155823)
- Joao Matos
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2017, Michel et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Functional mapping of yeast genomes by saturated transposition
eLife 6:e23570.
https://doi.org/10.7554/eLife.23570
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