| Yu Xinping,Ma Teng,Zhou Pingkun,Wu Yumei.Preliminary screening of novel IER5 interacting proteins after ionizing irradiation[J].Chinese Journal of Radiological Medicine and Protection,2017,37(4):246-250 |
| Preliminary screening of novel IER5 interacting proteins after ionizing irradiation |
| Received:November 11, 2016 |
| DOI：10.3760/cma.j.issn.0254-5098.2017.04.002 |
| KeyWords:Cervical cancer Immediate early response 5 Immuniprecipitation Mass spectrometry |
| FundProject:国家自然科学基金（81272888）；北京市医院管理局临床医学发展专项经费（ZYLX201705） |
| Author Name | Affiliation | E-mail | | Yu Xinping | Beijing Obstetric and Gynecological Hospital, Capital Medical University, Beijing 100006, China | | | Ma Teng | Institute of Radiation Medicine, the Military Medical Sciences, Beijing Key Laboratory for Radiobiology, Beijing 100850, China | | | Zhou Pingkun | Institute of Radiation Medicine, the Military Medical Sciences, Beijing Key Laboratory for Radiobiology, Beijing 100850, China | | | Wu Yumei | Beijing Obstetric and Gynecological Hospital, Capital Medical University, Beijing 100006, China | wym597118@163.com |
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| Abstract:: |
| Objective To investigate the expression of radiation-induced IER5 protein and screen its potential interaction proteins that may participate in DNA repair process. Methods HeLa cells were irradiated with 4 Gy ionizing radiation. IER5 protein expression in whole cell lysate and in nuclear fraction were detected by Western blot at different timepoints after irradiation. 3×Flag-IER5 pCMV plasmid was constrcuted and the Flag tagged-IER5 expression was verified by Western blot. 293T cells were transfected with 3×Flag-IER5 pCMV plasmid. After irradiation the cells were collected and proteins were extracted. The IER5 interaction proteins were purified using immunoprecipitation and separated by 12% SDS-polyacrylamide gel electrophoresis. Then the binding proteins were cut from the gel and analyzed by Mass spectrometry. Results The expression of IER5 protein began to increase 4 hour post-irradiation and its peak level was observed at 12 hour post-irradiation, and it lasted until 48 hour after irradiation. The expression level of IER5 protein in whole cell lysate and nuclear fraction were both increased. With the mass spectrometry analysis, a total of 347 proteins and 256 proteins were identified in irradiated and non-irradiated groups, respectively. Fourty one differential proteins were obtained, where 10 proteins were associated with DNA metabolic process and DNA rapair in the irradiated group and the poly(ADP-ribose) polymerase 1 (PARP1) protein was further confirmed by Western blot. Conclusions IER5 protein is an DNA damage related protein, and it may participate in DNA repair process. |
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