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18S and 25S Exonuclease Resistant Ribosomal RNA Molecules Are Produced by 5'-End Modification During TOR Inhibition.

Miguel A Rocha1, Gowda Bhavani1, Jacob Fleischmann1,2

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Yeast (Chichester, England)
|November 6, 2025
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Summary
This summary is machine-generated.

Saccharomyces cerevisiae yeast cells produce exonuclease-resistant ribosomal RNA (rRNA) during diauxic shift, linked to reduced TOR signaling. This resistance stems from 5' end modifications, not transcriptional changes.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Cellular Metabolism

Background:

  • Saccharomyces cerevisiae produces 18S and 25S ribosomal RNA (rRNA) resistant to exonuclease degradation.
  • This resistance is linked to the diauxic shift and reduced Target Of Rapamycin (TOR) signaling.
  • Exonuclease activity requires a 5' monophosphate, suggesting modifications to the rRNA 5' end confer resistance.

Purpose of the Study:

  • Investigate the relationship between TOR signaling and the accumulation of resistant rRNA.
  • Determine the mechanism and regulation of resistant rRNA formation.
  • Clarify the role of RNA polymerase I (Pol I) and posttranscriptional modifications.

Main Methods:

  • Pharmacological inhibition of TOR using rapamycin.
  • Analysis of resistant rRNA in TOR1-deleted mutants and constitutively active RNA polymerase I (CARA) cells.
  • Thiouracil labeling and uptake assays to assess rRNA synthesis and modification.
  • Decapping assays to evaluate exonuclease sensitivity.

Main Results:

  • Rapamycin treatment increased resistant 18S and 25S rRNA levels.
  • Resistant rRNA accumulation occurred independently of RNA Pol I transcriptional regulation (observed in CARA cells).
  • TOR1 deletion led to sustained production of resistant rRNA.
  • Log-phase rRNA can be converted to a resistant form post-transcriptionally.
  • Decapping restored exonuclease sensitivity, confirming 5' end modifications confer resistance.

Conclusions:

  • TOR signaling negatively regulates the accumulation of exonuclease-resistant rRNA.
  • Resistant rRNA formation is a posttranscriptional process involving 5' end modifications, likely phosphorylation.
  • This mechanism provides a novel layer of rRNA regulation during cellular stress or metabolic shifts.