Positioning of CCA-arms of the A- and the P-tRNAs towards the 28S rRNA in the human ribosome - Sorbonne Université
Article Dans Une Revue Biochimie Année : 2013

Positioning of CCA-arms of the A- and the P-tRNAs towards the 28S rRNA in the human ribosome

Konstantin Bulygin
  • Fonction : Auteur
Alexey Malygin
  • Fonction : Auteur
Codjo Hountondji
Dmitri Graifer
  • Fonction : Auteur
Galina Karpova
  • Fonction : Auteur
  • PersonId : 851986

Résumé

Nucleotides of 28S rRNA involved in binding of the human 80S ribosome with acceptor ends of the A site and the P site tRNAs were determined using two complementary approaches, namely, cross-linking with application of tRNAAsp analogues substituted with 4-thiouridine inposition 75 or 76 and hydroxyl radical footprinting with the use of the full sized tRNA and the tRNA deprived of the 30-terminal trinucleotide CCA. In general, these 28S rRNA nucleotides are located in ribosomal regions homologous to the A, P and E sites of the prokaryotic 50S subunit. However, none of the approaches used discovered interactions of the apex of the large rRNA helix 80 with the acceptor end of the P site tRNA typical with prokaryotic ribosomes. Application of the results obtained to available atomic models of 50S and 60S subunits led us to a conclusion that the A site tRNA is actually present in both A/A and A/P states and the P site tRNA in the P/P and P/E states. Thus, the present study gives a biochemical confirmation of the data on the structure and dynamics of the mammalian ribosomal pretranslocation complex obtained with application of cryo-electron microscopy and single-molecule FRET [Budkevich et al., 2011]. Moreover, in our study, particular sets of 28S rRNA nucleotides involved in oscillations of tRNAs CCA-termini between their alternative locations in the mammalian 80S ribosome are revealed.
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Dates et versions

hal-01906394 , version 1 (08-11-2018)

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Citer

Konstantin Bulygin, Alexey Malygin, Codjo Hountondji, Dmitri Graifer, Galina Karpova. Positioning of CCA-arms of the A- and the P-tRNAs towards the 28S rRNA in the human ribosome. Biochimie, 2013, 95 (2), pp.195 - 203. ⟨10.1016/j.biochi.2012.09.010⟩. ⟨hal-01906394⟩
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