Deficiency of the tRNA<sup>Tyr</sup>:Ψ35-synthase aPus7 in Archaea of the Sulfolobales order might be rescued by the H/ACA sRNA-guided machinery - Cancéropôle du Grand Est Accéder directement au contenu
Article Dans Une Revue Nucleic Acids Research Année : 2009

Deficiency of the tRNATyr:Ψ35-synthase aPus7 in Archaea of the Sulfolobales order might be rescued by the H/ACA sRNA-guided machinery

Résumé

Up to now, ψ formation in tRNAs was found to be catalysed by stand-alone enzymes. By computational analysis of archaeal genomes we detected putative H/ACA sRNAs, in four Sulfolobales species and in Aeropyrum pernix, that might guide ψ35 formation in pre-tRNATyr(GUA). This modification is achieved by Pus7p in eukarya. The validity of the computational predictions was verified by in vitro reconstitution of H/ACA sRNPs using the identified Sulfolobus solfataricus H/ACA sRNA. Comparison of Pus7-like enzymes encoded by archaeal genomes revealed amino acid substitutions in motifs IIIa and II in Sulfolobales and A. pernix Pus7-like enzymes. These conserved RNA:ψ-synthase-motifs are essential for catalysis. As expected, the recombinant Pyrococcus abyssi aPus7 was fully active and acted at positions 35 and 13 and other positions in tRNAs, while the recombinant S. solfataricus aPus7 was only found to have a poor activity at position 13. We showed that the presence of an A residue 3′ to the target U residue is required for P. abyssi aPus7 activity, and that this is not the case for the reconstituted S. solfataricus H/ACA sRNP. In agreement with the possible formation of ψ35 in tRNATyr(GUA) by aPus7 in P. abyssi and by an H/ACA sRNP in S. solfataricus, the A36G mutation in the P. abyssi tRNATyr(GUA) abolished ψ35 formation when using P. abyssi extract, whereas the A36G substitution in the S. solfataricus pre-tRNATyr did not affect ψ35 formation in this RNA when using an S. solfataricus extract. © 2009 The Author(s).
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hal-03992159 , version 1 (16-02-2023)

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Fabrice Leclerc, Christiane Branlant, Alan Urban, Sébastien Muller, Yuri Motorin, et al.. Deficiency of the tRNATyr:Ψ35-synthase aPus7 in Archaea of the Sulfolobales order might be rescued by the H/ACA sRNA-guided machinery. Nucleic Acids Research, 2009, 37 (4), pp.1308-1322. ⟨10.1093/nar/gkn1037⟩. ⟨hal-03992159⟩
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