rRNA pseudouridylation defects affect ribosomal ligand binding and translational fidelity from yeast to human cells

Research output: Contribution to journalArticle

Abstract

How pseudouridylation (Ψ), the most common and evolutionarily conserved modification of rRNA, regulates ribosome activity is poorly understood. Medically, Ψ is important because the rRNA Ψ synthase, DKC1, is mutated in X-linked dyskeratosis congenita (X-DC) and Hoyeraal-Hreidarsson (HH) syndrome. Here, we characterize ribosomes isolated from a yeast strain in which Cbf5p, the yeast homolog of DKC1, is catalytically impaired through a D95A mutation (cbf5-D95A). Ribosomes from cbf5-D95A cells display decreased affinities for tRNA binding to the A and P sites as well as the cricket paralysis virus internal ribosome entry site (IRES), which interacts with both the P and the E sites of the ribosome. This biochemical impairment in ribosome activity manifests as decreased translational fidelity and IRES-dependent translational initiation, which are also evident in mouse and human cells deficient for DKC1 activity. These findings uncover specific roles for Ψ modification in ribosome-ligand interactions that are conserved in yeast, mouse, and humans.

Details

Authors
  • Karen Jack
  • Cristian Bellodi
  • Dori M Landry
  • Rachel O Niederer
  • Arturas Meskauskas
  • Sharmishtha Musalgaonkar
  • Noam Kopmar
  • Olya Krasnykh
  • Alison M Dean
  • Sunnie R Thompson
  • Davide Ruggero
  • Jonathan D Dinman
Organisations
External organisations
  • University of California System
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Medical Genetics

Keywords

  • Animals, Binding Sites, Cell Cycle Proteins, Dyskeratosis Congenita, Fetal Growth Retardation, Genes, Reporter, Humans, Hydro-Lyases, Intellectual Disability, Luciferases, Mice, Microcephaly, Microtubule-Associated Proteins, Mutation, Nuclear Proteins, Plasmids, Protein Biosynthesis, RNA, Ribosomal, RNA, Transfer, Ribonucleoproteins, Small Nuclear, Ribosomes, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, Transduction, Genetic
Original languageEnglish
Pages (from-to)660-6
Number of pages7
JournalMolecular Cell
Volume44
Issue number4
Publication statusPublished - 2011 Nov 18
Publication categoryResearch
Peer-reviewedYes