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通过 mim-tRNAseq 对真核生物 tRNA 丰度和修饰状态进行高分辨率定量分析。

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

机构信息

Mechanisms of Protein Biogenesis, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.

Mechanisms of Protein Biogenesis, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany; Department of Chemistry, Technical University of Munich, 85748 Garching, Germany.

出版信息

Mol Cell. 2021 Apr 15;81(8):1802-1815.e7. doi: 10.1016/j.molcel.2021.01.028. Epub 2021 Feb 12.

DOI:10.1016/j.molcel.2021.01.028
PMID:33581077
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8062790/
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.

摘要

细胞 tRNA 丰度的测量受到修饰核苷普遍存在的 cDNA 合成障碍和 tRNA 基因之间广泛相似性的阻碍。我们通过修饰诱导的错配 tRNA 测序（mim-tRNAseq）克服了这些限制，该方法将从内源性修饰 tRNA 构建全长 cDNA 文库的工作流程与全面且用户友好的计算分析工具包相结合。我们的方法可以准确地捕获酵母、苍蝇和人类细胞中的 tRNA 丰度和修饰状态，并且适用于任何具有已知基因组的生物体。我们应用 mim-tRNAseq 来发现不同人类细胞系中 tRNA 同功密码子库的巨大异质性，以及同一 tRNA 转录本中不同位点的修饰之间令人惊讶的相互依赖性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/7b1f21b896fe/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/069ad9058781/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/9413dfccc665/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/46a529361070/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/65762229eef6/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/6bcb6e1c7c28/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/e868f8e5ce83/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/7b1f21b896fe/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/069ad9058781/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/9413dfccc665/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/46a529361070/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/65762229eef6/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/6bcb6e1c7c28/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/e868f8e5ce83/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/8062790/7b1f21b896fe/gr6.jpg

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