摘要
目的探讨RNA结合蛋白RBM45在DNA损伤应答中的功能。方法提取染色质结合蛋白后,Western印迹检测RBM45在DNA损伤位点的招募情况;利用Talen技术建立RBM45敲除的细胞系后,分别利用流式细胞仪和克隆形成实验检测敲除RBM45对细胞周期及细胞对DNA损伤敏感性的影响。结果DNA损伤后,RBM45可以迅速地被招募到DNA损伤位点;敲除RBM45后,Chk1磷酸化受抑制,细胞G2/M期检验点激活受阻;并且,敲除RBM45使得细胞对DNA损伤更敏感。结论RNA结合蛋白RBM45参与DNA损伤应答。
Objective To explore the role of RBM45in DNA damage response.Methods After eoUection of chromatin fractions,the recruitment of RBM45was detected by Western blot-ting.RBM45knockout (KO)cell line was generated by Talen technology.Cell cycle checkpoints and cell survival were determined in RBM45-KO cells by flowcytometry and clonogenic assay.Results RBM45was rapidly recruited to the chromatin after DNA damage.The phosphorylation of Chkl was decreased and the activation of G2/M checkpoint was delayed in RBM45-KO cells compared to that of WT cells.In addition,we found that knockout of RBM45sensitized the cells to irradiation killing.Conclusion The RNA binding protein RBM45is involved in DNA damage response.
作者
巩娟娟
刘红美
李巍
唐铁山
GONG Juanjuan;LIU Hongmei;LI Wei;TANG Tieshan(Beijing Children's Hospital,Capital Medical University,Beijing Pediatric Research Institute,Center for Medical Genetics,Beijing Key Laboratory for Genetics of Birth Defects,Beijing,100051,China;State Key Laboratory of Membrane Biology,Institute of Zoology,Beijing,100101,China)
出处
《医学分子生物学杂志》
CAS
2018年第6期369-372,共4页
Journal of Medical Molecular Biology
基金
北京市自然科学基金(No.5181001)。
