2. Academic Validation
  3. Genome-wide RNAi Screening Identifies Protein Modules Required for 40S Subunit Synthesis in Human Cells

Genome-wide RNAi Screening Identifies Protein Modules Required for 40S Subunit Synthesis in Human Cells

  • Cell Rep. 2015 Dec 29;13(12):2879-91. doi: 10.1016/j.celrep.2015.11.061.
Lukas Badertscher 1,  Thomas Wild 2,  Christian Montellese 1,  Leila T Alexander 3,  Lukas Bammert 1,  Marie Sarazova 1,  Michael Stebler 4,  Gabor Csucs 4,  Thomas U Mayer 5,  Nicola Zamboni 3,  Ivo Zemp 2,  Peter Horvath 6,  Ulrike Kutay 7
Affiliations

Affiliations

  • 1 Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; Molecular Life Sciences Ph.D. Program, 8057 Zurich, Switzerland.
  • 2 Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
  • 3 Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.
  • 4 RNAi Screening Center, ScopeM, ETH Zurich, 8093 Zurich, Switzerland.
  • 5 Department of Biology, University of Konstanz, 78457 Konstanz, Germany.
  • 6 Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; RNAi Screening Center, ScopeM, ETH Zurich, 8093 Zurich, Switzerland.
  • 7 Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland. Electronic address: ulrike.kutay@bc.biol.ethz.ch.
PMID: 26711351 DOI: 10.1016/j.celrep.2015.11.061
Abstract

Ribosome biogenesis is a highly complex process requiring many assisting factors. Studies in yeast have yielded comprehensive knowledge of the cellular machinery involved in this process. However, many aspects of ribosome synthesis are different in higher eukaryotes, and the global set of mammalian ribosome biogenesis factors remains unexplored. We used an imaging-based, genome-wide RNAi screen to find human proteins involved in 40S ribosomal subunit biogenesis. Our analysis identified ∼ 300 factors, many part of essential protein modules such as the small subunit (SSU) processome, the eIF3 and chaperonin complexes, and the ubiquitin-proteasome system. We demonstrate a role for the vertebrate-specific factor RBIS in ribosome synthesis, uncover a requirement for the CRL4 E3 ubiquitin Ligase in nucleolar ribosome biogenesis, and reveal that intracellular glutamine synthesis supports 40S subunit production.

Keywords

RNAi screening; cullin E3 ubiquitin ligase; glutamine metabolism; ribosome synthesis.

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