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Updated: Aug 5, 2025

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
09:04

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

Published on: July 26, 2018

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Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen

Daria S Spasskaya1, Kirill A Kulagin1,2, Evgenia N Grineva1,2

  • 1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia.

Journal of Fungi (Basel, Switzerland)
|March 29, 2023
PubMed

Insights

This study reveals that the proteasome directly regulates Ribonucleotide reductase (RNR), an enzyme crucial for DNA repair. Deregulation of the proteasome enhances yeast resistance to DNA damage.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Genomic stability is vital and maintained by DNA repair systems.
  • Ribonucleotide reductase (RNR) is essential for DNA repair by supplying deoxynucleotides.
  • Direct regulation of yeast RNR remains poorly understood.

Purpose of the Study:

  • To investigate the proteasome-dependent mechanism of direct RNR subunit regulation in Saccharomyces cerevisiae.
  • To understand the role of RNR in yeast resistance to DNA-damaging agents.

Main Methods:

  • Proteome analysis
  • Western blot
  • RT-PCR
  • Yeast plating analysis
  • Inhibition of RNR
  • Gene deletion studies

Main Results:

  • Proteasome deregulation leads to RNR upregulation.
  • Upregulated RNR confers hyper-resistance to 4-nitroquinoline-1-oxide (4-NQO).
  • Inhibition of RNR or deletion of its regulatory proteins abolishes 4-NQO resistance.
  • Yeast Rnr1 subunit is identified as a proteasome substrate.

Conclusions:

  • The proteasome directly regulates RNR activity in yeast.
  • This regulation impacts cellular resistance to DNA damage.
  • Findings suggest a conserved RNR regulation mechanism between yeast and mammals.
Keywords:
4-NQOS. cerevisiaeproteasomeribonucleotide reductaseshotgun proteomics

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