RNF8 promotes efficient DSB repair by inhibiting the pro‐apoptotic activity of p53 through regulating the function of Tip60
| Autor: | Jialing Liu, Yunpeng Feng, Yueshuang Ke, Hongyu Chen, Xianlu Zeng, Wenjing Qi, Jin Shan, Wenguang Liu |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
p53
0301 basic medicine DNA Repair Ubiquitin-Protein Ligases Cell Apoptosis Lysine Acetyltransferase 5 RNF8 Flow cytometry law.invention 03 medical and health sciences 0302 clinical medicine Western blot law Cell Line Tumor medicine Humans DNA Breaks Double-Stranded Gene knockdown cell apoptosis medicine.diagnostic_test biology Cell growth Chemistry DNA double‐strand break (DSB) repair Original Articles Cell Biology General Medicine HCT116 Cells Cell biology Ubiquitin ligase DNA-Binding Proteins cell proliferation 030104 developmental biology medicine.anatomical_structure Tip60 030220 oncology & carcinogenesis biology.protein Suppressor Original Article Tumor Suppressor Protein p53 |
| Zdroj: | Cell Proliferation |
| ISSN: | 1365-2184 0960-7722 |
| DOI: | 10.1111/cpr.12780 |
| Popis: | Objectives RING finger protein 8 (RNF8) is an E3 ligase that plays an essential role in DSB repair. p53 is a well‐established tumour suppressor and cellular gatekeeper of genome stability. This study aimed at investigating the functional correlations between RNF8 and p53 in DSB damage repair. Materials and methods In this article, wild‐type, knockout and shRNA‐depleted HCT116 and U2OS cells were stressed, and the roles of RNF8 and p53 were examined. RT‐PCR and Western blot were utilized to investigate the expression of related genes in damaged cells. Cell proliferation, apoptosis and neutral cell comet assays were applied to determine the effects of DSB damage on differently treated cells. DR‐GFP, EJ5‐GFP and LacI‐LacO targeting systems, flow cytometry, mass spectrometry, IP, IF, GST pull‐down assay were used to explore the molecular mechanism of RNF8 and p53 in DSB damage repair. Results We found that RNF8 knockdown increased cellular sensitivity to DSB damage and decreased cell proliferation, which was correlated with high expression of the p53 gene. RNF8 improved the efficiency of DSB repair by inhibiting the pro‐apoptotic function of p53. We also found that RNF8 restrains cell apoptosis by inhibiting over‐activation of ATM and subsequently reducing p53 acetylation at K120 through regulating Tip60. Conclusions Taken together, these findings suggested that RNF8 promotes efficient DSB repair by inhibiting the pro‐apoptotic activity of p53 through regulating the function of Tip60. |
| Databáze: | OpenAIRE |
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