The deubiquitinating enzyme USP37 enhances CHK1 activity to promote the cellular response to replication stress
| Autor: | Andrew Dickson, Benjamin R. Stromberg, Christine Insinna, Matthew K. Summers, Yosup Rhee, Adrian E. Torres, Debjani Pal, Christopher J. Westlake, Mayank Singh, Amy C. Burrows |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
SCFβTRCP
SKP1–CUL1–F-box in complex with the β-transducing repeat–containing protein DNA damage response Biochemistry Deubiquitinating enzyme S Phase Histones Enzyme Stability HDAC6 histone deacetylase 6 checkpoint control deubiquitination Ub ubiquitin βTRCP β-transducing repeat–containing protein biology Chemistry Cell cycle Cell biology USP37 DUB deubiquitinating enzyme ATR ataxia telangiectasia and Rad3-related MCF-7 Cells cell cycle PBST PBS with Tween-20 Research Article DNA Replication CHK1 CHK1 checkpoint kinase 1 ubiquitination APH aphidicolin DDR DNA damage response Genomic Instability CHX cycloheximide APC/CCDH1 cyclosome in complex with the activator CDH1 GST glutathione-S-transferase Endopeptidases Humans CHEK1 APC/C anaphase-promoting complex/Cyclosome TBST Tris-buffered saline with Tween-20 Molecular Biology Mitosis Cell growth DNA replication Cell Biology SCF SKP1–CUL1–F-box HDAC6 HCT116 Cells Checkpoint Kinase 1 biology.protein BSA bovine serum albumin Ataxia telangiectasia and Rad3 related HU hydroxyurea DNA Damage HeLa Cells |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | The deubiquitinating enzyme USP37 is known to contribute to timely onset of S phase and progression of mitosis. However, it is not clear if USP37 is required beyond S-phase entry despite expression and activity of USP37 peaking within S phase. We have utilized flow cytometry and microscopy to analyze populations of replicating cells labeled with thymidine analogs and monitored mitotic entry in synchronized cells to determine that USP37-depleted cells exhibited altered S-phase kinetics. Further analysis revealed that cells depleted of USP37 harbored increased levels of the replication stress and DNA damage markers γH2AX and 53BP1 in response to perturbed replication. Depletion of USP37 also reduced cellular proliferation and led to increased sensitivity to agents that induce replication stress. Underlying the increased sensitivity, we found that the checkpoint kinase 1 is destabilized in the absence of USP37, attenuating its function. We further demonstrated that USP37 deubiquitinates checkpoint kinase 1, promoting its stability. Together, our results establish that USP37 is required beyond S-phase entry to promote the efficiency and fidelity of replication. These data further define the role of USP37 in the regulation of cell proliferation and contribute to an evolving understanding of USP37 as a multifaceted regulator of genome stability. |
| Databáze: | OpenAIRE |
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