Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic>
ABSTRACT Protein ubiquitylation regulates not only endocellular trafficking and proteasomal degradation but also the catalytic activity of enzymes. In Saccharomyces cerevisiae, we analyzed the composition of the ubiquitylated proteomes in strains lacking acetyltransferase Gcn5p, Ub-protease Ubp8p, o...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2020-08-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.01504-20 |
| _version_ | 1819203122199789568 |
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| author | Antonella De Palma Giulia Fanelli Elisabetta Cretella Veronica De Luca Raffaele Antonio Palladino Valentina Panzeri Valentina Roffia Michele Saliola Pierluigi Mauri Patrizia Filetici |
| author_facet | Antonella De Palma Giulia Fanelli Elisabetta Cretella Veronica De Luca Raffaele Antonio Palladino Valentina Panzeri Valentina Roffia Michele Saliola Pierluigi Mauri Patrizia Filetici |
| author_sort | Antonella De Palma |
| collection | DOAJ |
| description | ABSTRACT Protein ubiquitylation regulates not only endocellular trafficking and proteasomal degradation but also the catalytic activity of enzymes. In Saccharomyces cerevisiae, we analyzed the composition of the ubiquitylated proteomes in strains lacking acetyltransferase Gcn5p, Ub-protease Ubp8p, or both to understand their involvement in the regulation of protein ubiquitylation. We analyzed His6Ub proteins with a proteomic approach coupling micro-liquid chromatography and tandem mass spectrometry (μLC-MS/MS) in gcn5Δ, ubp8Δ and ubp8Δ gcn5Δ strains. The Ub-proteome altered in the absence of Gcn5p, Ubp8p, or both was characterized, showing that 43% of the proteins was shared in all strains, suggesting their functional relationship. Remarkably, all major glycolytic enzymes showed increased ubiquitylation. Phosphofructokinase 1, the key enzyme of glycolytic flux, showed a higher and altered pattern of ubiquitylation in gcn5Δ and ubp8Δ strains. Severe defects of growth in poor sugar and altered glucose consumption confirmed a direct role of Gcn5p and Ubp8p in affecting the REDOX balance of the cell. IMPORTANCE We propose a study showing a novel role of Gcn5p and Ubp8p in the process of ubiquitylation of the yeast proteome which includes main glycolytic enzymes. Interestingly, in the absence of Gcn5p and Ubp8p glucose consumption and redox balance were altered in yeast. We believe that these results and the role of Gcn5p and Ubp8p in sugar metabolism might open new perspectives of research leading to novel protocols for counteracting the enhanced glycolysis in tumors. |
| first_indexed | 2024-12-23T04:14:53Z |
| format | Article |
| id | doaj.art-bd81e473ee064372a1520e4593a9d3b9 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-23T04:14:53Z |
| publishDate | 2020-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-bd81e473ee064372a1520e4593a9d3b92022-12-21T18:00:24ZengAmerican Society for MicrobiologymBio2150-75112020-08-0111410.1128/mBio.01504-20Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic>Antonella De Palma0Giulia Fanelli1Elisabetta Cretella2Veronica De Luca3Raffaele Antonio Palladino4Valentina Panzeri5Valentina Roffia6Michele Saliola7Pierluigi Mauri8Patrizia Filetici9Proteomics and Metabolomics Unit, Institute for Biomedical Technologies (ITB-CNR), Segrate, ItalyInstitute of Molecular Biology and Pathology/CNR, Sapienza University of Rome, Rome, ItalyInstitute of Molecular Biology and Pathology/CNR, Sapienza University of Rome, Rome, ItalyInstitute of Molecular Biology and Pathology/CNR, Sapienza University of Rome, Rome, ItalyInstitute of Molecular Biology and Pathology/CNR, Sapienza University of Rome, Rome, ItalyProteomics and Metabolomics Unit, Institute for Biomedical Technologies (ITB-CNR), Segrate, ItalyProteomics and Metabolomics Unit, Institute for Biomedical Technologies (ITB-CNR), Segrate, ItalyDepartment of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, Rome, ItalyProteomics and Metabolomics Unit, Institute for Biomedical Technologies (ITB-CNR), Segrate, ItalyInstitute of Molecular Biology and Pathology/CNR, Sapienza University of Rome, Rome, ItalyABSTRACT Protein ubiquitylation regulates not only endocellular trafficking and proteasomal degradation but also the catalytic activity of enzymes. In Saccharomyces cerevisiae, we analyzed the composition of the ubiquitylated proteomes in strains lacking acetyltransferase Gcn5p, Ub-protease Ubp8p, or both to understand their involvement in the regulation of protein ubiquitylation. We analyzed His6Ub proteins with a proteomic approach coupling micro-liquid chromatography and tandem mass spectrometry (μLC-MS/MS) in gcn5Δ, ubp8Δ and ubp8Δ gcn5Δ strains. The Ub-proteome altered in the absence of Gcn5p, Ubp8p, or both was characterized, showing that 43% of the proteins was shared in all strains, suggesting their functional relationship. Remarkably, all major glycolytic enzymes showed increased ubiquitylation. Phosphofructokinase 1, the key enzyme of glycolytic flux, showed a higher and altered pattern of ubiquitylation in gcn5Δ and ubp8Δ strains. Severe defects of growth in poor sugar and altered glucose consumption confirmed a direct role of Gcn5p and Ubp8p in affecting the REDOX balance of the cell. IMPORTANCE We propose a study showing a novel role of Gcn5p and Ubp8p in the process of ubiquitylation of the yeast proteome which includes main glycolytic enzymes. Interestingly, in the absence of Gcn5p and Ubp8p glucose consumption and redox balance were altered in yeast. We believe that these results and the role of Gcn5p and Ubp8p in sugar metabolism might open new perspectives of research leading to novel protocols for counteracting the enhanced glycolysis in tumors.https://journals.asm.org/doi/10.1128/mBio.01504-20ubiquitylationGcn5pUbp8pglycolytic fluxsugar utilization |
| spellingShingle | Antonella De Palma Giulia Fanelli Elisabetta Cretella Veronica De Luca Raffaele Antonio Palladino Valentina Panzeri Valentina Roffia Michele Saliola Pierluigi Mauri Patrizia Filetici Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic> mBio ubiquitylation Gcn5p Ubp8p glycolytic flux sugar utilization |
| title | Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic> |
| title_full | Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic> |
| title_fullStr | Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic> |
| title_full_unstemmed | Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic> |
| title_short | Gcn5p and Ubp8p Affect Protein Ubiquitylation and Cell Proliferation by Altering the Fermentative/Respiratory Flux Balance in <italic toggle="yes">Saccharomyces cerevisiae</italic> |
| title_sort | gcn5p and ubp8p affect protein ubiquitylation and cell proliferation by altering the fermentative respiratory flux balance in italic toggle yes saccharomyces cerevisiae italic |
| topic | ubiquitylation Gcn5p Ubp8p glycolytic flux sugar utilization |
| url | https://journals.asm.org/doi/10.1128/mBio.01504-20 |
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