title: Genome-wide RNAi Screening Identifies Protein Modules Required for 40S Subunit Synthesis in Human Cells.
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.
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; Molecular Life Sciences Ph.D. Program, 8057 Zurich, Switzerland.
authors.forename: Lukas
authors.initials: L
authors.lastname: Badertscher
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wild
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; Molecular Life Sciences Ph.D. Program, 8057 Zurich, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Montellese
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Leila T
authors.initials: LT
authors.lastname: Alexander
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; Molecular Life Sciences Ph.D. Program, 8057 Zurich, Switzerland.
authors.forename: Lukas
authors.initials: L
authors.lastname: Bammert
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; Molecular Life Sciences Ph.D. Program, 8057 Zurich, Switzerland.
authors.forename: Marie
authors.initials: M
authors.lastname: Sarazova
authors.affiliation: RNAi Screening Center, ScopeM, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Stebler
authors.affiliation: RNAi Screening Center, ScopeM, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Gabor
authors.initials: G
authors.lastname: Csucs
authors.affiliation: Department of Biology, University of Konstanz, 78457 Konstanz, Germany.
authors.forename: Thomas U
authors.initials: TU
authors.lastname: Mayer
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Nicola
authors.initials: N
authors.lastname: Zamboni
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Ivo
authors.initials: I
authors.lastname: Zemp
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland; RNAi Screening Center, ScopeM, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Horvath
authors.affiliation: Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland. Electronic address: ulrike.kutay@bc.biol.ethz.ch.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Kutay
mesh_terms: D023281:Genomics / Q000379:methods*; D005973:Glutamine / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D034622:RNA Interference; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000096:biosynthesis* / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D053722:Receptors, Interleukin-17
chemical_list: D005973:Glutamine; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; D044767:Ubiquitin-Protein Ligases; C408922:IL17RB protein, human; D053722:Receptors, Interleukin-17
keywords: RNAi screening; cullin E3 ubiquitin ligase; glutamine metabolism; ribosome synthesis