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. Epub 2015 Dec 17.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Genomics / methods*
  • Glutamine / metabolism
  • HeLa Cells
  • Humans
  • RNA Interference*
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism*
  • Ribosomal Proteins / biosynthesis*
  • Ribosomal Proteins / genetics
  • Ubiquitin-Protein Ligases / metabolism


  • IL17RB protein, human
  • RNA, Ribosomal
  • Ribosomal Proteins
  • Glutamine
  • Ubiquitin-Protein Ligases