Journal
NUCLEIC ACIDS RESEARCH
Volume 48, Issue 11, Pages 6210-6222Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkaa315
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Funding
- Swiss National Science Foundation
- NCCR RNA Disease
- ETH Zurich from European Research Council [EURIBIO260676]
- Olga Mayenfisch Stiftung
- Spanish Ministry of Economy and Competitiveness (MINECO)
- European Union ERFD program [BFU2012-32404, BFU2016-75352-P]
- Spanish Ministry of Education
- EMBO short-stay fellowship
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The ribotoxin alpha-sarcin belongs to a family of ribonucleases that cleave the sarcin/ricin loop (SRL), a critical functional rRNA element within the large ribosomal subunit (60S), thereby abolishing translation. Whether alpha-sarcin targets the SRL only in mature 60S subunits remains unresolved. Here, we show that, in yeast, alpha-sarcin can cleave SRLs within late 60S pre-ribosomes containing mature 25S rRNA but not nucleolar/nuclear 60S pre-ribosomes containing 27S pre-rRNA in vivo. Conditional expression of alpha-sarcin is lethal, but does not impede early pre-rRNA processing, nuclear export and the cytoplasmic maturation of 60S pre-ribosomes. Thus, SRL-cleaved containing late 60S pre-ribosomes seem to escape cytoplasmic proofreading steps. Polysome analyses revealed that SRL-cleaved 60S ribosomal subunits form 80S initiation complexes, but fail to progress to the step of translation elongation. We suggest that the functional integrity of a alpha-sarcin cleaved SRL might be assessed only during translation.
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