组蛋白甲基转移酶SETD2对P53介导细胞凋亡的调控  

Histone methyltransferase SETD2 in regulation of P53-mediated apoptosis

作　　者：谢萍[1,2] 田春艳[2] 张令强[2] 贺福初[1,2] 

机构地区：[1]清华大学生命科学学院,北京100084 [2]军事医学科学院放射与辐射医学研究所北京蛋白质组研究中心蛋白质组学国家重点实验室,北京100850

出　　处：《军事医学》2011年第1期30-34,共5页Military Medical Sciences

基　　金：国家自然科学基金(30770518);国家973计划项目(2007CB914601)

摘　　要：目的探讨组蛋白甲基转移酶SETD2对P53功能的调控。方法 PCR扩增人源SETD2并重组入真核表达载体;通过间接免疫荧光鉴定SETD2的亚细胞定位;通过GST Pull-down实验寻找P53结合SETD2的区域;流式细胞术分析SETD2对P53参与细胞凋亡的影响。结果成功克隆了人源SETD2的全长,并将其成功构建到真核表达载体中;SETD2以胞核定位为主;体外直接相互作用实验证实SETD2和P53存在相互作用,SETD2分子中部的SET结构域和C端都可以结合P53;流式细胞分析结果表明SETD2对于P53参与的细胞凋亡具有明显的促进作用。结论本研究揭示P53正向调控因子SETD2对P53介导的细胞凋亡具有促进效应,从而将组蛋白表遗传学修饰与P53转录因子活性调控关联起来,研究结果对肿瘤、免疫等疾病的防治具有一定的科学意义。Objective To investigate the role of the histone methyltransferase SETD2 on the regulation of P53-mediated apoptosis.Methods Amplified the Setd2 gene by PCR to recombine it into the eukaryotic expression vector.Using indirect immunofluorescence assay（IIF） to analyze the subcellular localization of Myc-tagged SETD2 protein.A glutathione S-transferase（GST） pull-down assay was performed to detect the direct interaction between P53 and SETD2 truncates.Flow cytometry（FCM） was used to study the effect of SETD2 on P53-dependent cell apoptosis.Results Setd2 gene was amplified by PCR and constructed into eukaryotic expression vector.SETD2 protein was mainly localized in the nucleus.GST pull-down assay showed that SETD2 interacted with P53 directly.The SET domain and the C-terminus of SETD2 mediated the interaction with P53.Overexpression of SETD2 enhanced the P53-promoted cell apoptosis.Conclusion These findings suggest that the histone methyltransferase SETD2 upregulate P53-dependent apoptosis.

关 键 词：蛋白甲基转移酶类 SETD2 肿瘤抑制蛋白质P53 聚合酶链反应 真核表达 细胞凋亡 流式细胞术 

分 类 号：Q255[生物学—细胞生物学]