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

Jack, Karen; Bellodi, Cristian LU ; Landry, Dori M; Niederer, Rachel O; Meskauskas, Arturas; Musalgaonkar, Sharmishtha; Kopmar, Noam; Krasnykh, Olya; Dean, Alison M and Thompson, Sunnie R, et al. (2011) In Molecular Cell 44(4). p.6-660
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... (More)

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.

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@misc{21155f13-a460-4755-bb6d-7925968b8683,
  abstract     = {<p>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.</p>},
  author       = {Jack, Karen and Bellodi, Cristian and Landry, Dori M and Niederer, Rachel O and Meskauskas, Arturas and Musalgaonkar, Sharmishtha and Kopmar, Noam and Krasnykh, Olya and Dean, Alison M and Thompson, Sunnie R and Ruggero, Davide and Dinman, Jonathan D},
  issn         = {1097-4164},
  keyword      = {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},
  language     = {eng},
  month        = {11},
  number       = {4},
  pages        = {6--660},
  publisher    = {ARRAY(0x8e644c8)},
  series       = {Molecular Cell},
  title        = {rRNA pseudouridylation defects affect ribosomal ligand binding and translational fidelity from yeast to human cells},
  url          = {http://dx.doi.org/10.1016/j.molcel.2011.09.017},
  volume       = {44},
  year         = {2011},
}