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High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq.

ABSTRACT: Measurements of cellular tRNA abundance are hampered by pervasive blocks to cDNA synthesis at modified nucleosides and the extensive similarity among tRNA genes. We overcome these limitations with modification-induced misincorporation tRNA sequencing (mim-tRNAseq), which combines a workflow for full-length cDNA library construction from endogenously modified tRNA with a comprehensive and user-friendly computational analysis toolkit. Our method accurately captures tRNA abundance and modification status in yeast, fly, and human cells and is applicable to any organism with a known genome. We applied mim-tRNAseq to discover a dramatic heterogeneity of tRNA isodecoder pools among diverse human cell lines and a surprising interdependence of modifications at distinct sites within the same tRNA transcript.

SUBMITTER: Behrens A

PROVIDER: S-EPMC8062790 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq.

Behrens Andrew A Rodschinka Geraldine G Nedialkova Danny D DD

Molecular cell 20210212 8

Measurements of cellular tRNA abundance are hampered by pervasive blocks to cDNA synthesis at modified nucleosides and the extensive similarity among tRNA genes. We overcome these limitations with modification-induced misincorporation tRNA sequencing (mim-tRNAseq), which combines a workflow for full-length cDNA library construction from endogenously modified tRNA with a comprehensive and user-friendly computational analysis toolkit. Our method accurately captures tRNA abundance and modification ...[more]

PMID: 33581077

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