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 and Lionel Benard ()
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Albertas Navickas: Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes
Sébastien Chamois: Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes
Rénette Saint-Fort: Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes
Julien Henri: Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes
Claire Torchet: Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes
Lionel Benard: Laboratoire de Biologie moléculaire et Cellulaire des Eucaryotes

Nature Communications, 2020, vol. 11, issue 1, 1-11

Abstract: 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. Here we use 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.

Date: 2020
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DOI: 10.1038/s41467-019-13991-9

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