Functional mapping of yeast genomes by saturated transposition

  1. Agnès Henria Michel
  2. Riko Hatakeyama
  3. Philipp Kimmig
  4. Meret Arter
  5. Matthias Peter
  6. Joao Matos
  7. Claudio De Virgilio
  8. Benoît Kornmann  Is a corresponding author
  1. ETH Zurich, Switzerland
  2. 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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Author details

  1. Agnès Henria Michel

    Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  2. Riko Hatakeyama

    Department of Biology, University of Fribourg, Fribourg, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  3. Philipp Kimmig

    Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  4. Meret Arter

    Institute of Biochemistry, ETH Zurich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  5. Matthias Peter

    Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2160-6824
  6. Joao Matos

    Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3754-3709
  7. Claudio De Virgilio

    Department of Biology, University of Fribourg, Fribourg, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  8. Benoît Kornmann

    Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
    For correspondence
    benoit.kornmann@bc.biol.ethz.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6030-8555

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.

Reviewing Editor

  1. Raymond J Deshaies, California Institute of Technology, United States

Publication history

  1. Received: November 23, 2016
  2. Accepted: May 6, 2017
  3. Accepted Manuscript published: May 8, 2017 (version 1)
  4. Version of Record published: June 9, 2017 (version 2)
  5. Version of Record updated: October 26, 2017 (version 3)

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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  1. Agnès Henria Michel
  2. Riko Hatakeyama
  3. Philipp Kimmig
  4. Meret Arter
  5. Matthias Peter
  6. Joao Matos
  7. Claudio De Virgilio
  8. Benoît Kornmann
(2017)
Functional mapping of yeast genomes by saturated transposition
eLife 6:e23570.
https://doi.org/10.7554/eLife.23570

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