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Formation of the conserved pseudouridine at position 55 in archaeal tRNA
Roovers, M.; Hale, C.; Tricot, C.; Terns, M.P.; Terns, R.M.; Grosjean, H.; Droogmans, L. (2006). Formation of the conserved pseudouridine at position 55 in archaeal tRNA. Nucleic Acids Res. 34(15): 4293-4301. https://dx.doi.org/10.1093/nar/gkl530
In: Nucleic Acids Research. Information Retrieval: London. ISSN 0305-1048; e-ISSN 1362-4962, more
Peer reviewed article  

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  • Roovers, M.
  • Hale, C.
  • Tricot, C.
  • Terns, M.P.
  • Terns, R.M.
  • Grosjean, H.
  • Droogmans, L., more

Abstract
    Pseudouridine (Ψ) located at position 55 in tRNA is a nearly universally conserved RNA modification found in all three domains of life. This modification is catalyzed by TruB in bacteria and by Pus4 in eukaryotes, but so far the Ψ55 synthase has not been identified in archaea. In this work, we report the ability of two distinct pseudouridine synthases from the hyperthermophilic archaeon Pyrococcus furiosus to specifically modify U55 in tRNA in vitro. These enzymes are pfuCbf5, a protein known to play a role in RNA-guided modification of rRNA, and pfuPsuX, a previously uncharacterized enzyme that is not a member of the TruB/Pus4/Cbf5 family of pseudouridine synthases. pfuPsuX is hereafter renamed pfuPus10. Both enzymes specifically modify tRNA U55 in vitro but exhibit differences in substrate recognition. In addition, we find that in a heterologous in vivo system, pfuPus10 efficiently complements an Escherichia coli strain deficient in the bacterial Ψ55 synthase TruB. These results indicate that it is probable that pfuCbf5 or pfuPus10 (or both) is responsible for the introduction of pseudouridine at U55 in tRNAs in archaea. While we cannot unequivocally assign the function from our results, both possibilities represent unexpected functions of these proteins as discussed herein.

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