Differential Regulation of p53 function by Lysine Acetylation

赖氨酸乙酰化对 p53 功能的差异调节

• 批准号: 7257003
• 负责人: SYED Shiraz MUJTABA
• 金额: $ 8.23万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-06 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7257003
• 关键词: Acetylation; Address; Amino Acids; Apoptosis; BAX gene; Bax protein; Binding; Biological Assay; Biological Process; Bromodomain; C-terminal; CDKN1A gene; CREB-binding protein; Camptothecin; Cell Cycle Arrest; Cell physiology; Cells; Chemicals; Complex; Condition; Consensus; Data; Doxorubicin/Etoposide; EP300 gene; Event; Figs - dietary; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Growth; HCT116 Cells; Histone Code; Histones; Hour; Human; Individual; Kinetics; Link; Lysine; MDM2 gene; MDM2 gene; Malignant Neoplasms; Mediating; Methylnitronitrosoguanidine; Modification; Molecular; Mutation; N-terminal; PAC1 phosphatase; PCAF gene; Pattern; Phosphorylation; Physiological; Play; Post-Translational Protein Processing; Protein p53; Proteins; Publishing; Radiation; Reagent; Regulation; Reporting; Role; Serine; Site; Stress; TP53 gene; Tail; Transcription Coactivator; Transcriptional Activation; Tumor Cell Line; Tumor Suppressor Proteins; UV induced; Ubiquitination; base; cell growth; chromatin immunoprecipitation; combinatorial; human CREBBP protein; inhibitor/antagonist; knock-down; oncoprotein p21; outcome forecast; response; senescence; size; small molecule; transcription factor; tumor; Acetylation; Address; Amino Acids; Apoptosis; BAX gene; Bax protein; Binding; Biological Assay; Biological Process; Bromodomain; C-terminal; CDKN1A gene; CREB-binding protein; Camptothecin; Cell Cycle Arrest; Cell physiology; Cells; Chemicals; Complex; Condition; Consensus; Data; Doxorubicin/Etoposide; EP300 gene; Event; Figs - dietary; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Growth; HCT116 Cells; Histone Code; Histones; Hour; Human; Individual; Kinetics; Link; Lysine; MDM2 gene; MDM2 gene; Malignant Neoplasms; Mediating; Methylnitronitrosoguanidine; Modification; Molecular; Mutation; N-terminal; PAC1 phosphatase; PCAF gene; Pattern; Phosphorylation; Physiological; Play; Post-Translational Protein Processing; Protein p53; Proteins; Publishing; Radiation; Reagent; Regulation; Reporting; Role; Serine; Site; Stress; TP53 gene; Tail; Transcription Coactivator; Transcriptional Activation; Tumor Cell Line; Tumor Suppressor Proteins; UV induced; Ubiquitination; base; cell growth; chromatin immunoprecipitation; combinatorial; human CREBBP protein; inhibitor/antagonist; knock-down; oncoprotein p21; outcome forecast; response; senescence; size; small molecule; transcription factor; tumor

DESCRIPTION (provided by applicant): Stress-induced C-terminal lysine acetylation of p53 plays an important role in the activity of p53 as a transcription factor that regulates cell cycle arrest, senescence or apoptosis. The long-term goal of this Project is to seek mechanistic understanding of the molecular interactions that regulate the tumor suppressor p53. While multiple acetylation sites in its C-terminal tail have been reported, specific effects of individual or combined acetylation of these lysine residues on p53 activity remain elusive. Preliminary data is presented involving the analysis of p53 function that acetylation-induced p53 activation in response to DMA damage is involved in co-activator recruitment. This study revealed that p53 recruitment of the co-activator CBP (CREB binding protein) requires association of the conserved bromodomain of CBP with p53 at acetylated Iys382: a specific molecular interaction that is essential for p53-induced transcriptional activation of the cyclin- dependent kinase inhibitor p21, involved in G1 cell cycle arrest. We hypothesize that distinct modifications of p53 C-terminal residues, including lysine acetylation and ubiquitination, as well as serine phosphorylation, have differential effects on p53 functions in cells under different conditions. A multifaceted approach is proposed to address mechanistic underpinnings of p53 transcriptional activation with the emphasis on the role of C-terminal post-translational modifications in p53 activation. To understand the complex molecular mechanisms underlying p53 regulation, I propose to investigate the dynamics of acetylation and molecular interactions of p53 involving its C-terminal lysines. Three specific aims are proposed to achieve this goal: (1) to investigate the kinetics of acetylation of the C-terminal lysines in p53 function in normal and tumor cell lines; (2) to elucidate the differential roles of the co-activators p300 and CBP in p53 regulation using CHIP and siRNAs; and (3) to modulate p53 activity between cell growth arrest and apoptosis using the newly developed CBP Bromodomain-binding small molecule inhibitors. The emerging results from the proposed studies are expected to enhance our understanding of the molecular basis of C-terminal modifications in p53 function. Given the central role of p53 in cancer, these studies will have important implications for the prognosis and treatment of human tumors.

描述（由申请人提供）：p53的应力诱导的C末端赖氨酸乙酰化在p53的活性中起着重要的作用，作为调节细胞周期停滞，衰老或凋亡的转录因子。该项目的长期目标是寻求对调节肿瘤抑制p53的分子相互作用的机械理解。虽然已经报道了其C末端尾部中的多个乙酰化位点，但这些赖氨酸残基对p53活性的个体或乙酰化的特定影响仍然难以捉摸。提出了初步数据，其中涉及p53功能的分析，即乙酰化诱导的p53激活对DMA损伤涉及共激活因子募集。这项研究表明，p53共激活因子CBP（CREB结合蛋白）募集需要将CBP保守的溴结构域与乙酰化Iys382：一种特定的分子相互作用相关联，这是p53诱导的依赖性基因酶的p53诱导的p5依赖性kinase celtor p21 In libiby p21的特定分子相互作用。我们假设p53 C末端残基（包括赖氨酸乙酰化和泛素化）以及丝氨酸磷酸化的明显修饰对在不同条件下细胞中p53功能具有不同的影响。提出了一种多方面的方法来解决p53转录激活的机械基础，重点是C末端翻译后修饰在p53激活中的作用。为了了解p53调节的基础的复杂分子机制，我建议研究乙酰化和涉及其C末端赖氨酸的乙酰化和分子相互作用的动力学。提出了三个具体目的以实现这一目标：（1）研究正常和肿瘤细胞系中p53功能中C末端赖氨酸的乙酰化动力学； （2）使用芯片和siRNA阐明了共激活因子p300和cbp在p53调节中的差异作用； （3）使用新开发的CBP溴结构域结合小分子抑制剂来调节细胞生长停滞和凋亡之间的p53活性。预计拟议研究的新兴结果将增强我们对p53功能中C末端修饰的分子基础的理解。鉴于p53在癌症中的核心作用，这些研究将对人类肿瘤的预后和治疗具有重要意义。