Ubiquitin and Not Only Unfolded Domains Drives Toscana Virus Non-Structural NSs Protein Degradation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Viruses
2.2. Reagents and Antibodies
2.3. Plasmids
2.4. Cycloheximide Chase Analysis and NSs Protein Stability
2.5. Pull-Down and Immunoblot Analysis
2.6. Mass Spectrometry Detection of NSs Ubiquitination
3. Results
3.1. NSs Stability Is Influenced by Disordered Regions
3.2. Ubiquitin-Dependent NSs Proteasomal Degradation
3.3. Evidence of TOSV NSs Ubiquitination
3.4. Specific NSs Lysine Residues Undergo Ubiquitination
4. Discussion
Author Contributions
Funding
Conflicts of Interest
References
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Lysine Position | Percentage Sequence Coverage | Peptide Sequence | Peptide Identification Probability | Mascot Ion Score | Mascot Identity Score | Mascot Delta Ion Score | Variable Modifications Identified by Spectrum |
---|---|---|---|---|---|---|---|
180 | 7.21% | VLIEGKkHGLTAFDLPGNDILGDICVVQAAR | 99.70% | 58.6 | 60.3 | 36.67597403 | K7:GlyGly (+114.04) |
180 | 7.21% | VLIEGKkHGLTAFDLPGNDILGDICVVQAAR | 99.70% | 45.9 | 33.3 | 32.14675325 | K7:GlyGly (+114.04) |
180 | 6.94% | kHGLTAFDLPGNDILGDICVVQAAR | 99.70% | 63.2 | 48.6 | 55.33246753 | K1:GlyGly (+114.04) |
244 | 3.80% | KEDk | 99.70% | 55.4 | 60 | 21.31764706 | K4:GlyGly (+114.04) |
244 | 6.33% | KEDkRAKAKGLmSmCAAR | 99.70% | 51.2 | 43.7 | 31.05 | K4:GlyGly (+114.04) |
244 | 6.33% | EDkRAKAKGLMSMCAAR | 99.70% | 48.7 | 50.4 | 30.65454545 | K3:GlyGly (+114.04) |
294–298 | 6.59% | TDLGFRETALSTFWAKDWPTPQETILSDkRcLkEDMR | 99.70% | 48.6 | 36 | 34.75324675 | K29:GlyGly (+114.04) K33:GlyGly (+114.04) |
294–298 | 6.33% | DWPTLQETILSDkRcLkEDmRVTK | 99.70% | 52.6 | 38 | 43.26406926 | K13:GlyGly (+114.04) K17:GlyGly (+114.04) |
294 | 6.33% | ETALSTFWAKDWPTPQETILSDk | 99.70% | 60.9 | 65.5 | 23.27176471 | K23:GlyGly (+114.04) |
298 | 6.33% | CLkEDMRVTKWLPSPPHYPPL | 99.70% | 45.8 | 38.3 | 27.21315789 | K4:GlyGly (+114.04) |
298 | 6.33% | CLKEDMRVTKWLPSPPHYPPL | 99.70% | 44.2 | 45.9 | 27.91753247 | K23:GlyGly (+114.04) |
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Gori Savellini, G.; Bini, L.; Gagliardi, A.; Anichini, G.; Gandolfo, C.; Prathyumnan, S.; Cusi, M.G. Ubiquitin and Not Only Unfolded Domains Drives Toscana Virus Non-Structural NSs Protein Degradation. Viruses 2020, 12, 1153. https://doi.org/10.3390/v12101153
Gori Savellini G, Bini L, Gagliardi A, Anichini G, Gandolfo C, Prathyumnan S, Cusi MG. Ubiquitin and Not Only Unfolded Domains Drives Toscana Virus Non-Structural NSs Protein Degradation. Viruses. 2020; 12(10):1153. https://doi.org/10.3390/v12101153
Chicago/Turabian StyleGori Savellini, Gianni, Luca Bini, Assunta Gagliardi, Gabriele Anichini, Claudia Gandolfo, Shibily Prathyumnan, and Maria Grazia Cusi. 2020. "Ubiquitin and Not Only Unfolded Domains Drives Toscana Virus Non-Structural NSs Protein Degradation" Viruses 12, no. 10: 1153. https://doi.org/10.3390/v12101153