USP29 Deubiquitinates SETD8 and Regulates DNA Damage-Induced H4K20 Monomethylation and 53BP1 Focus Formation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Treatments
2.2. siRNA Oligos, Plasmids, and Transfection
GFP | CAAGCUGACCCUGAAGUUCdTdT |
Luc | CGUACGCGGAAUACUUCGAdTdT |
SETD8 | GUACGGAGCGCCAUGAAGUdTdT |
USP29#1 | CCCAUCAAGUUUAGAGGAUdTdT |
USP29#2 | GGAAUAUGCUGAAGGAAAUdTdT |
TRABID | AGAGGCUUCUUCAAUAAUAdTdT |
USP18 | CCAGGGAGUUAUCAAGCAAdTdT |
USP2 | CAGAUUGUGGUUACUGUUCUAdTdT |
USP30 | CCAGAGUCCUGUUCGAUUUdTdT |
USP33 | UCUCGACAGUGGCUUAAUUAAdTdT |
USP51 | CCAUUUAGCUGUAGACCUUdTdT |
2.3. Antibodies
2.4. Immunoprecipitations, Pulldowns, and Western Blot
2.5. Immunofluorescence
2.6. Flow Cytometry
2.7. Clonogenic Survival
2.8. Statistical Analysis and Reproducibility
3. Results
3.1. USP29 Controls SETD8 Protein Levels
3.2. USP29 Regulates SETD8 Levels in a Cell-Cycle-Independent Manner
3.3. SETD8 and USP29 Interact In Vivo
3.4. USP29 Deubiquitinates SETD8 In Vivo
3.5. Accumulation of 53BP1 into Nuclear Foci upon IR Is Regulated by USP29
3.6. USP29 Controls IR-Induced H4K20 Monomethylation and Cell Survival
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Hernández-Reyes, Y.; Paz-Cabrera, M.C.; Freire, R.; Smits, V.A.J. USP29 Deubiquitinates SETD8 and Regulates DNA Damage-Induced H4K20 Monomethylation and 53BP1 Focus Formation. Cells 2022, 11, 2492. https://doi.org/10.3390/cells11162492
Hernández-Reyes Y, Paz-Cabrera MC, Freire R, Smits VAJ. USP29 Deubiquitinates SETD8 and Regulates DNA Damage-Induced H4K20 Monomethylation and 53BP1 Focus Formation. Cells. 2022; 11(16):2492. https://doi.org/10.3390/cells11162492
Chicago/Turabian StyleHernández-Reyes, Yeray, María Cristina Paz-Cabrera, Raimundo Freire, and Veronique A. J. Smits. 2022. "USP29 Deubiquitinates SETD8 and Regulates DNA Damage-Induced H4K20 Monomethylation and 53BP1 Focus Formation" Cells 11, no. 16: 2492. https://doi.org/10.3390/cells11162492