The endonuclease cue2 cleaves mRNAs at stalled ribosomes during no go decay

Karole N. D’Orazio, Colin Chih Chien Wu, Niladri Sinha, Raphael Loll-Krippleber, Grant W. Brown, Rachel Green

Research output: Contribution to journalArticle

Abstract

Translation of problematic sequences in mRNAs leads to ribosome collisions that trigger a series of quality control events including ribosome rescue, degradation of the stalled nascent polypeptide, and targeting of the mRNA for decay (No Go Decay or NGD). Using a reverse genetic screen in yeast, we identify Cue2 as the conserved endonuclease that is recruited to stalled ribosomes to promote NGD. Ribosome profiling and biochemistry provide strong evidence that Cue2 cleaves mRNA within the A site of the colliding ribosome. We demonstrate that NGD primarily proceeds via Xrn1-mediated exonucleolytic decay and Cue2-mediated endonucleolytic decay normally constitutes a secondary decay pathway. Finally, we show that the Cue2-dependent pathway becomes a major contributor to NGD in cells depleted of factors required for the resolution of stalled ribosome complexes. Together these results provide insights into how multiple decay processes converge to process problematic mRNAs in eukaryotic cells.

Original languageEnglish (US)
Article numbere49117
JournaleLife
Volume8
DOIs
StatePublished - Jun 2019

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Endonucleases
Ribosomes
Messenger RNA
Biochemistry
Yeast
Quality control
Reverse Genetics
RNA Stability
Eukaryotic Cells
Degradation
Quality Control
Peptides
Yeasts

ASJC Scopus subject areas

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

D’Orazio, K. N., Wu, C. C. C., Sinha, N., Loll-Krippleber, R., Brown, G. W., & Green, R. (2019). The endonuclease cue2 cleaves mRNAs at stalled ribosomes during no go decay. eLife, 8, [e49117]. https://doi.org/10.7554/eLife.49117.001

The endonuclease cue2 cleaves mRNAs at stalled ribosomes during no go decay. / D’Orazio, Karole N.; Wu, Colin Chih Chien; Sinha, Niladri; Loll-Krippleber, Raphael; Brown, Grant W.; Green, Rachel.

In: eLife, Vol. 8, e49117, 06.2019.

Research output: Contribution to journalArticle

D’Orazio, KN, Wu, CCC, Sinha, N, Loll-Krippleber, R, Brown, GW & Green, R 2019, 'The endonuclease cue2 cleaves mRNAs at stalled ribosomes during no go decay', eLife, vol. 8, e49117. https://doi.org/10.7554/eLife.49117.001
D’Orazio, Karole N. ; Wu, Colin Chih Chien ; Sinha, Niladri ; Loll-Krippleber, Raphael ; Brown, Grant W. ; Green, Rachel. / The endonuclease cue2 cleaves mRNAs at stalled ribosomes during no go decay. In: eLife. 2019 ; Vol. 8.
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