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Nanopore direct RNA sequencing confirms coordinated base modifications in the T-loop of S. cerevisiae tRNA


Three nucleoside modifications in the tRNA T-loop are conserved across life: 5-methyluridine (m5U), pseudouridine (ψ), and N1-methyladenosine (m1A).

  • T-loop modifying enzymes in S. cerevisiae:
    • m5U catalyzed by trm2
    • ψ-55 catalyzed by pus4
    • m1A catalyzed by gcd10/gcd14

In S. cerevisiae their addition is coordinated in a modification circuit, as has been demonstrated using NMR spectroscopy (Barraud 2019; Yared 2022).

Nanopore direct RNA sequencing is capable of detecting certain modifications like ψ and m1A.

We applied Nanopore direct RNA sequencing to investigate this modification circuit in all 42 S. cerevisiae tRNA isoacceptors.

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