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