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No-Go Decay mRNA cleavage in the ribosome exit tunnel produces 5’-OH ends phosphorylated by Trl1

Albertas Navickas, Sébastien Chamois, Rénette Saint-Fort, Julien Henri, Claire Torchet, Lionel Benard
doi: https://doi.org/10.1101/465633
Albertas Navickas
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes, Paris, France
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Sébastien Chamois
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes, Paris, France
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Rénette Saint-Fort
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes, Paris, France
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Julien Henri
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes, Paris, France
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Claire Torchet
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes, Paris, France
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Lionel Benard
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes, Paris, France
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  • For correspondence: lionel.benard@ibpc.fr
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Abstract

The No-Go Decay (NGD) mRNA surveillance pathway degrades mRNAs containing stacks of stalled ribosomes. Although an endoribonuclease has been proposed to initiate cleavages upstream of the stall sequence, the production of two RNA fragments resulting from a unique cleavage has never been demonstrated. We have used mRNAs expressing a 3’-ribozyme to produce truncated transcripts in vivo to mimic naturally occurring truncated mRNAs known to trigger NGD. This technique allows us to analyse endonucleolytic cleavage events at single-nucleotide resolution starting at the third collided ribosome, which we show to be Hel2-dependent. These cleavages map precisely in the mRNA exit tunnel of the ribosome, 8 nucleotides upstream of the first P-site residue and release 5’-hydroxylated RNA fragments requiring 5’-phosphorylation prior to digestion by the exoribonuclease Xrn1, or alternatively by Dxo1. Finally, we identify the RNA kinase Trl1, alias Rlg1, as an essential player in the degradation of NGD RNAs.

Footnotes

  • Updated version with detailed information in the Methods section and clarified Figures 1f and 5b

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Posted December 09, 2019.
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No-Go Decay mRNA cleavage in the ribosome exit tunnel produces 5’-OH ends phosphorylated by Trl1
Albertas Navickas, Sébastien Chamois, Rénette Saint-Fort, Julien Henri, Claire Torchet, Lionel Benard
bioRxiv 465633; doi: https://doi.org/10.1101/465633
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No-Go Decay mRNA cleavage in the ribosome exit tunnel produces 5’-OH ends phosphorylated by Trl1
Albertas Navickas, Sébastien Chamois, Rénette Saint-Fort, Julien Henri, Claire Torchet, Lionel Benard
bioRxiv 465633; doi: https://doi.org/10.1101/465633

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