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人类 NMD 独立于稳定的核糖体停滞而发生。

Human NMD ensues independently of stable ribosome stalling.

机构信息

Department of Chemistry and Biochemistry, University of Bern, CH-3012, Bern, Switzerland.

Graduate School for Cellular and Biomedical Sciences, University of Bern, CH-3012, Bern, Switzerland.

出版信息

Nat Commun. 2020 Aug 17;11(1):4134. doi: 10.1038/s41467-020-17974-z.


DOI:10.1038/s41467-020-17974-z
PMID:32807779
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7431590/
Abstract

Nonsense-mediated mRNA decay (NMD) is a translation-dependent RNA degradation pathway that is important for the elimination of faulty, and the regulation of normal, mRNAs. The molecular details of the early steps in NMD are not fully understood but previous work suggests that NMD activation occurs as a consequence of ribosome stalling at the termination codon (TC). To test this hypothesis, we established an in vitro translation-coupled toeprinting assay based on lysates from human cells that allows monitoring of ribosome occupancy at the TC of reporter mRNAs. In contrast to the prevailing NMD model, our in vitro system reveals similar ribosomal occupancy at the stop codons of NMD-sensitive and NMD-insensitive reporter mRNAs. Moreover, ribosome profiling reveals a similar density of ribosomes at the TC of endogenous NMD-sensitive and NMD-insensitive mRNAs in vivo. Together, these data show that NMD activation is not accompanied by stable stalling of ribosomes at TCs.

摘要

无意义介导的 mRNA 降解 (NMD) 是一种依赖翻译的 RNA 降解途径,对于消除错误的和调节正常的 mRNA 非常重要。NMD 早期步骤的分子细节尚未完全了解,但先前的工作表明,NMD 的激活是核糖体在终止密码子 (TC) 处停滞的结果。为了验证这一假说,我们建立了一种基于人细胞裂解物的体外翻译偶联的足迹分析测定法,该方法允许监测报告 mRNA 上 TC 处的核糖体占有率。与流行的 NMD 模型相反,我们的体外系统显示 NMD 敏感和 NMD 不敏感报告 mRNA 的 TC 处的核糖体占有率相似。此外,核糖体谱分析显示,体内 NMD 敏感和 NMD 不敏感的内源性 mRNA 的 TC 处的核糖体密度相似。总之,这些数据表明 NMD 的激活并不伴随着核糖体在 TC 处的稳定停滞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/ae990d53c83a/41467_2020_17974_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/ae255d4a2c8f/41467_2020_17974_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/a7f10bd1c798/41467_2020_17974_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/be606154f633/41467_2020_17974_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/809ea802ef55/41467_2020_17974_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/ae990d53c83a/41467_2020_17974_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/ae255d4a2c8f/41467_2020_17974_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/a7f10bd1c798/41467_2020_17974_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/be606154f633/41467_2020_17974_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/809ea802ef55/41467_2020_17974_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0f/7431590/ae990d53c83a/41467_2020_17974_Fig5_HTML.jpg

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Human NMD ensues independently of stable ribosome stalling.

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[4]
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[5]
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[6]
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[7]
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[8]
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[9]
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[10]
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引用本文的文献

[1]
Efficient cell-free translation from diverse human cell types.

J Biol Chem. 2025-5-28

[2]
Cytoplasmic mRNA decay and quality control machineries in eukaryotes.

Nat Rev Genet. 2025-1-27

[3]
Mechanisms and Delivery of tRNA Therapeutics.

Chem Rev. 2024-6-26

[4]
Comprehensive translational profiling and STE AI uncover rapid control of protein biosynthesis during cell stress.

Nucleic Acids Res. 2024-7-22

[5]
Nonsense-mediated mRNA decay pathway in plants under stress: general gene regulatory mechanism and advances.

Planta. 2024-1-30

[6]
Universal features of Nsp1-mediated translational shutdown by coronaviruses.

Mol Cell. 2023-10-5

[7]
Translation-coupled mRNA quality control mechanisms.

EMBO J. 2023-10-4

[8]
Universal features of Nsp1-mediated translational shutdown by coronaviruses.

bioRxiv. 2023-6-1

[9]
Engineered tRNAs suppress nonsense mutations in cells and in vivo.

Nature. 2023-6

[10]
Coupled protein quality control during nonsense-mediated mRNA decay.

J Cell Sci. 2023-5-15

本文引用的文献

[1]
Cell-free gene expression: an expanded repertoire of applications.

Nat Rev Genet. 2019-11-28

[2]
Nonsense-Mediated mRNA Decay in Development, Stress and Cancer.

Adv Exp Med Biol. 2019

[3]
Single-Molecule Imaging Uncovers Rules Governing Nonsense-Mediated mRNA Decay.

Mol Cell. 2019-5-30

[4]
Nonsense-mediated mRNA decay: The challenge of telling right from wrong in a complex transcriptome.

Wiley Interdiscip Rev RNA. 2019-5-26

[5]
Quality and quantity control of gene expression by nonsense-mediated mRNA decay.

Nat Rev Mol Cell Biol. 2019-7

[6]
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RNA. 2019-1-17

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Nonsense-Mediated mRNA Decay Begins Where Translation Ends.

Cold Spring Harb Perspect Biol. 2019-2-1

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Roadblocks and resolutions in eukaryotic translation.

Nat Rev Mol Cell Biol. 2018-8

[9]
Highly efficient in vitro translation of authentic affinity-purified messenger ribonucleoprotein complexes.

RNA. 2018-5-3

[10]
Dual function of UPF3B in early and late translation termination.

EMBO J. 2017-10-16

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