title: rRNA pseudouridylation defects affect ribosomal ligand binding and translational fidelity from yeast to human cells.
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.
authors.affiliation: Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Jack
authors.forename: Cristian
authors.initials: C
authors.lastname: Bellodi
authors.forename: Dori M
authors.initials: DM
authors.lastname: Landry
authors.forename: Rachel O
authors.initials: RO
authors.lastname: Niederer
authors.forename: Arturas
authors.initials: A
authors.lastname: Meskauskas
authors.forename: Sharmishtha
authors.initials: S
authors.lastname: Musalgaonkar
authors.forename: Noam
authors.initials: N
authors.lastname: Kopmar
authors.forename: Olya
authors.initials: O
authors.lastname: Krasnykh
authors.forename: Alison M
authors.initials: AM
authors.lastname: Dean
authors.forename: Sunnie R
authors.initials: SR
authors.lastname: Thompson
authors.forename: Davide
authors.initials: D
authors.lastname: Ruggero
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Dinman
mesh_terms: D000818:Animals; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000172:deficiency* / Q000235:genetics; D019871:Dyskeratosis Congenita / Q000201:enzymology / Q000235:genetics*; D005317:Fetal Growth Retardation / Q000201:enzymology / Q000235:genetics*; D017930:Genes, Reporter; D006801:Humans; D006836:Hydro-Lyases / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D008607:Intellectual Disability / Q000201:enzymology / Q000235:genetics*; D008156:Luciferases / Q000032:analysis; D051379:Mice; D008831:Microcephaly / Q000201:enzymology / Q000235:genetics*; D008869:Microtubule-Associated Proteins / Q000172:deficiency* / Q000235:genetics; D009154:Mutation; D009687:Nuclear Proteins / Q000172:deficiency* / Q000235:genetics; D010957:Plasmids; D014176:Protein Biosynthesis; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear / Q000172:deficiency* / Q000235:genetics; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014161:Transduction, Genetic
chemical_list: D018797:Cell Cycle Proteins; C112148:DKC1 protein, human; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D012335:RNA, Ribosomal; D017411:Ribonucleoproteins, Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer; D008156:Luciferases; D006836:Hydro-Lyases; C082278:CBF5 protein, S cerevisiae; C022479:pseudouridylate synthetase
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