A Novel Role for RNF126 in the Promotion of G2 Arrest via Interaction With 14-3-3σ
| Autor: | Qi-En Wang, Junran Zhang, Zhaojun Qiu, Chunhong Yan, Pengyan Fa |
|---|---|
| Rok vydání: | 2022 |
| Předmět: |
Cancer Research
Cell cycle checkpoint DNA repair Article chemistry.chemical_compound Radiation Ionizing Medicine Radiology Nuclear Medicine and imaging Propidium iodide CHEK1 Cyclin B1 Radiation biology business.industry Kinase Cell Cycle Checkpoints Cell cycle Ubiquitin ligase Cell biology G2 Phase Cell Cycle Checkpoints Oncology chemistry biology.protein M Phase Cell Cycle Checkpoints business DNA Damage |
| Zdroj: | Int J Radiat Oncol Biol Phys |
| ISSN: | 0360-3016 |
| DOI: | 10.1016/j.ijrobp.2021.09.025 |
| Popis: | Purpose Cell cycle checkpoints and DNA repair are important for cell survival after exogenous DNA damage. Both rapid blockage of G2 to M phase transition in the cell cycle and the maintenance of relatively slow G2 arrest are critical to protect cells from lethal ionizing radiation (IR). Checkpoint kinase 1 is pivotal in blocking the transition from G2 to M phases in response to IR. The 14-3-3σ protein is important for IR-induced G2 arrest maintenance in which p53-dependent 14-3-3σ transcription is involved. It has been demonstrated that Ring finger protein 126 (RNF126), an E3 ligase, is required to upregulate checkpoint kinase 1 expression. Thus, our goal was to study the role of RNF126 in the G2/M phase checkpoint. Methods and Materials The transition from G2 to M phases and G2 accumulation in response to IR were determined by flow cytometry through staining with phospho-histone H3 (pS10) antibody and propidium iodide, respectively. The interaction of RNF126 and 14-3-3σ was determined by GST-pulldown and coimmunoprecipitation assays. The stability of RNF126 and 14-3-3σ was determined by cycloheximide-based stability assay and ubiquitination detection by coimmunoprecipitation. The sequestering of cyclin-dependent kinase 1 and cyclin B1 from the nucleus was determined by immunofluorescence staining. Results RNF126 knockdown had no impact on the IR-induced transient blockage of G2 to M but impaired IR-induced G2 arrest maintenance in cells with or without wild-type p53. Mechanistically, RNF126 binds 14-3-3σ and prevents both proteins from ubiquitination-mediated degradation. Last, RNF126 is required for enforcing the cytoplasmic sequestration of cyclin B1 and cyclin-dependent kinase 1 proteins in response to IR. Conclusions RNF126 promotes G2 arrest via interaction with 14-3-3σ in response to IR. Our study revealed a novel role for RNF126 in promoting G2 arrest, providing a new target for cancer treatment. |
| Databáze: | OpenAIRE |
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