摘要
肿瘤抑制蛋白p53作为一种重要的转录因子,参与细胞周期调节、细胞凋亡、细胞衰老以及DNA修复等生物学过程。p53在不同生理条件下发挥相应的功能离不开众多辅因子的协助,这些辅因子可以调节p53的修饰状态、细胞亚定位,以及p53的稳定性等。因此,鉴定p53结合蛋白质对进一步理解p53在体内的信号调控网络具有重要生物学意义。本文利用酵母双杂交实验技术筛选出1个新的p53结合蛋白质,FADD样白细胞介素1β转化酶相关巨蛋白(FADD-like interleukin-1β-converting enzyme associated huge protein,FLASH)。免疫共沉淀分析证实了FLASH与p53之间存在相互作用,并进一步阐明了这两个蛋白质之间相互作用的结构域基础。p53可同时与FLASH-N1(aa1~200)和FLASH-C1(aa1534~1780)相互作用。此外,FLASH-N、FLASH-C都能与p53-C(aa293~393)相互作用。通过报告基因研究FLASH对p53转录活性的影响,结果表明,FLASH全长以及FLASH-M(aa921~1533)能够增强p53的转录活性。综上所述,FLASH能够结合p53并增强其转录活性。
As an important transcription factor,the tumor suppressor protein p53 is involved in multiple biological processes such as cell cycle regulation,cell division,cell senescence and DNA repair.The functional roles of p53 under various physiological conditions are inseparable from the assistance of many cofactors,which can regulate the protein modification,cellular sub-localization,and protein stability of p53.Therefore,the identification of p53-binding protein(s)has important biological significance for further understanding the signal transduction network of p53 in vivo.In the present study,a novel p53-binding protein,FADD-like interleukin-1β-converting enzyme associated huge protein(FLASH)was identified through a yeast two-hybrid screen.The protein-protein interaction and the structural basis of the interaction between FLASH and p53 was also confirmed by co-immunoprecipitation analysis.These studies have shown that p53 can simultaneously interact with both FLASH-N1(aa 1~200)and FLASH-C1(aa 1534~1780).In addition,both of FLASH-N and FLASH-C can interact with the same region of p53(aa 293~393).Transcription analysis has revealed that the full length of FLASH and FLASH-M(aa 921~1533)can enhance the transcription activity of p53.In summary,FLASH can bind to p53 and enhance its transcriptional activity.
作者
陈小雁
熊真真
尤姗姗
石晓钟
CHEN Xiao-Yan;XIONG Zhen-Zhen;YOU Shan-Shan;SHI Xiao-Zhong(School of Biology and Biological Engineering,Department of Biotechnology,South China University of Technology,Guangzhou 510006,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2021年第10期1345-1356,共12页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金项目(No.31771449)资助。
