Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO
Various external and internal factors damaging DNA constantly disrupt the stability of the genome. Cells use numerous dedicated DNA repair systems to detect damage and restore genomic integrity in a timely manner. Ribonucleotide reductase (RNR) is a key enzyme providing dNTPs for DNA repair. Molecul...
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| Format: | Article |
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MDPI AG
2023-03-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/9/3/351 |
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| author | Daria S. Spasskaya Kirill A. Kulagin Evgenia N. Grineva Pamila J. Osipova Svetlana V. Poddubko Julia A. Bubis Elizaveta M. Kazakova Tomiris T. Kusainova Vladimir A. Gorshkov Frank Kjeldsen Vadim L. Karpov Irina A. Tarasova Dmitry S. Karpov |
| author_facet | Daria S. Spasskaya Kirill A. Kulagin Evgenia N. Grineva Pamila J. Osipova Svetlana V. Poddubko Julia A. Bubis Elizaveta M. Kazakova Tomiris T. Kusainova Vladimir A. Gorshkov Frank Kjeldsen Vadim L. Karpov Irina A. Tarasova Dmitry S. Karpov |
| author_sort | Daria S. Spasskaya |
| collection | DOAJ |
| description | Various external and internal factors damaging DNA constantly disrupt the stability of the genome. Cells use numerous dedicated DNA repair systems to detect damage and restore genomic integrity in a timely manner. Ribonucleotide reductase (RNR) is a key enzyme providing dNTPs for DNA repair. Molecular mechanisms of indirect regulation of yeast RNR activity are well understood, whereas little is known about its direct regulation. The study was aimed at elucidation of the proteasome-dependent mechanism of direct regulation of RNR subunits in <i>Saccharomyces cerevisiae</i>. Proteome analysis followed by Western blot, RT-PCR, and yeast plating analysis showed that upregulation of RNR by proteasome deregulation is associated with yeast hyper resistance to 4-nitroquinoline-1-oxide (4-NQO), a UV-mimetic DNA-damaging drug used in animal models to study oncogenesis. Inhibition of RNR or deletion of RNR regulatory proteins reverses the phenotype of yeast hyper resistance to 4-NQO. We have shown for the first time that the yeast Rnr1 subunit is a substrate of the proteasome, which suggests a common mechanism of RNR regulation in yeast and mammals. |
| first_indexed | 2024-03-11T06:19:52Z |
| format | Article |
| id | doaj.art-1f72b77b8ce941f2bf18f6e03c2c68ae |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-11T06:19:52Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-1f72b77b8ce941f2bf18f6e03c2c68ae2023-11-17T12:00:21ZengMDPI AGJournal of Fungi2309-608X2023-03-019335110.3390/jof9030351Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQODaria S. Spasskaya0Kirill A. Kulagin1Evgenia N. Grineva2Pamila J. Osipova3Svetlana V. Poddubko4Julia A. Bubis5Elizaveta M. Kazakova6Tomiris T. Kusainova7Vladimir A. Gorshkov8Frank Kjeldsen9Vadim L. Karpov10Irina A. Tarasova11Dmitry S. Karpov12Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, RussiaInstitute of Medical and Biological Problems, Russian Academy of Sciences, 123007 Moscow, RussiaInstitute of Medical and Biological Problems, Russian Academy of Sciences, 123007 Moscow, RussiaV.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences, 119334 Moscow, RussiaV.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences, 119334 Moscow, RussiaV.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences, 119334 Moscow, RussiaDepartment of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, DenmarkDepartment of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, DenmarkEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, RussiaV.L. Talrose Institute for Energy Problems of Chemical Physics, N.N. Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences, 119334 Moscow, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, RussiaVarious external and internal factors damaging DNA constantly disrupt the stability of the genome. Cells use numerous dedicated DNA repair systems to detect damage and restore genomic integrity in a timely manner. Ribonucleotide reductase (RNR) is a key enzyme providing dNTPs for DNA repair. Molecular mechanisms of indirect regulation of yeast RNR activity are well understood, whereas little is known about its direct regulation. The study was aimed at elucidation of the proteasome-dependent mechanism of direct regulation of RNR subunits in <i>Saccharomyces cerevisiae</i>. Proteome analysis followed by Western blot, RT-PCR, and yeast plating analysis showed that upregulation of RNR by proteasome deregulation is associated with yeast hyper resistance to 4-nitroquinoline-1-oxide (4-NQO), a UV-mimetic DNA-damaging drug used in animal models to study oncogenesis. Inhibition of RNR or deletion of RNR regulatory proteins reverses the phenotype of yeast hyper resistance to 4-NQO. We have shown for the first time that the yeast Rnr1 subunit is a substrate of the proteasome, which suggests a common mechanism of RNR regulation in yeast and mammals.https://www.mdpi.com/2309-608X/9/3/351shotgun proteomics4-NQOribonucleotide reductase<i>S. cerevisiae</i>proteasome |
| spellingShingle | Daria S. Spasskaya Kirill A. Kulagin Evgenia N. Grineva Pamila J. Osipova Svetlana V. Poddubko Julia A. Bubis Elizaveta M. Kazakova Tomiris T. Kusainova Vladimir A. Gorshkov Frank Kjeldsen Vadim L. Karpov Irina A. Tarasova Dmitry S. Karpov Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO Journal of Fungi shotgun proteomics 4-NQO ribonucleotide reductase <i>S. cerevisiae</i> proteasome |
| title | Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO |
| title_full | Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO |
| title_fullStr | Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO |
| title_full_unstemmed | Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO |
| title_short | Yeast Ribonucleotide Reductase Is a Direct Target of the Proteasome and Provides Hyper Resistance to the Carcinogen 4-NQO |
| title_sort | yeast ribonucleotide reductase is a direct target of the proteasome and provides hyper resistance to the carcinogen 4 nqo |
| topic | shotgun proteomics 4-NQO ribonucleotide reductase <i>S. cerevisiae</i> proteasome |
| url | https://www.mdpi.com/2309-608X/9/3/351 |
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