Purα蛋白质调控p53蛋白乙酰化的研究分析论文.docVIP

中国科技论文在线   Purα蛋白质调控 p53 蛋白乙酰化的研究 吴怡，章小斌，贾军祥，张风群，丁明，赵辅昆** （浙江理工大学生命科学学院，杭州 310018） 5 10 15 20 25 30 摘要：蛋白质翻译后修饰包括有甲基化、乙酰化、磷酸化和泛素化等。这些修饰可改变染色 体的状态，影响转录因子与 DNA 序列的结合，能对基因的表达起调控作用。Purα是一个多 功能蛋白，广泛存在于生物体内且能与特定 DNA 和 RNA 核酸序列结合，Purα通过启动 DNA 复制、调控转录，从而参与 mRNA 运输与翻译以及 DNA 损伤修复的过程。而体内 p53 蛋白参 与 DNA 损伤修复，调控细胞增殖与凋亡，参与细胞分化与转录调节。实验发现 Purα和 p53 都具有参与 DNA 损伤的功能。本文构建了 pIRES-Flag-Purα重组表达载体，通过转染人胚 肾 293T 细胞，筛选获得了稳定表达 Purα的 293T 细胞株。Werstern Blot 实验结果表明 Pur α表达能够抑制 p53 蛋白 373 位赖氨酸的乙酰化。细胞免疫荧光定位实验表明 Purα在 DNA 损伤情况下有入核现象。以上这些实验工作为进一步研究 Purα参与 DNA 损伤修复的机制奠 定了基础。 关键词：Purα；p53；DNA 损伤 中图分类号：Q513 Purα Protein can regulate the p53 K373 Acetylation level Wu Yi, Zhang Xiaobing, Jia Junxiang, Zhang Fengqun, Ding Ming, Zhao Fukun (School of Life Science,Zhejiang Sci-Tech University,Hangzhou 310018) Abstract: Protein modifications are including methylation, acethlation, phosphorylation, ubiquitination and so on. Most of these modifications can change the status of chromosomes, affect the binding of transcription factor with DNA sequence, and regulate gene expression by several mechanisms. Purα is a multifunctional and ubiquitous nucleic acid-binding protein that can bind to both DNA. Purα takes part in the transport and translation of mRNA and the damage repair of DNA by starting DNA replication and transcription regulation.In vivo, p53 exerts a pivotal role in controlling the apoptosis, cell cycle and DNA repair in response to various forms of genotoxic stress. We found that Purα and p53 both carry with the DNA Damage. We constructed pIRES-Flag-Purα vector, and got the stable 293T cell lines of expression Purα.Search the impact of Purα to p53, we found over express Purα that suppress acetylation of p53 at lysine 373.We found that Purα from cytoplasm into nuclear phenomena under the DNA damage conditions through immunofluorescence. The above works have provided a foundation for further research mechanism of Purα in repair of DNA damage. Key words: Purα; p53; DNA damage 35 0 引言 Purα 是一种广泛存在于