Structural basis for CBP/p300 transcriptional regulation

CBP/p300 转录调控的结构基础

• 批准号: 7909484
• 负责人: PETER Edwin WRIGHT
• 金额: $ 5.55万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7909484
• 关键词: Acetylation; Address; Adenovirus E1A Proteins; Affect; Affinity; Apoptosis; Binding; Binding Sites; Biological Assay; CREB-binding protein; Cell Cycle Regulation; Cell physiology; Cellular Stress; Collaborations; Complex; DNA Repair; Data; Differentiation and Growth; EP300 gene; Embryonic Development; Eukaryotic Cell; Event; Genetic Transcription; Genotoxic Stress; Goals; Growth; Individual; Infection; Libraries; MDM2 gene; Measures; Mediating; Modeling; Molecular; Multiple Partners; N-terminal; Normal Cell; Nuclear; Oncogene Proteins; Oncogenic; Oncogenic Viruses; Phosphorylation; Play; Proliferating; Property; Protein p53; Proteins; Research; Retinoblastoma Protein; Roentgen Rays; Role; Serotyping; Signal Transduction Pathway; Site; Structure; TP53 gene; Tertiary Protein Structure; Transactivation; Transcription Coactivator; Transcriptional Regulation; Tumor Suppression; Tumor Suppressor Proteins; Viral; Viral Oncogene; base; cell immortalization; human CREBBP protein; immortalized cell; in vitro testing; in vivo; insight; molecular recognition; non-oncogenic; repaired; research study; response; retinoblastoma tumor suppressor; transcription factor

DESCRIPTION (provided by applicant): CREB binding protein (CBP) and p300 are modular transcriptional coactivators that integrate numerous signal transduction pathways in eukaryotic cells, and are essential for growth, differentiation, apoptosis, and DNA repair. CBP/p300 play critical roles in embryonic development and are the targets of many oncogenic events. They also function as tumor suppressors and are required for the transcriptional activities of many cellular and tumor virus oncoproteins. CBP and p300 play an essential role in mediating the stability and transcriptional activity of the tumor suppressor p53. The interactions between these proteins are highly dynamic and are modulated by phosphorylation of the p53 transactivation domain at multiple sites in response to genotoxic stress. The adenoviral oncoprotein E1A competes with cellular factors to sequester CBP/p300, disrupting normal cell cycle control to induce cell immortalization. This research will utilize an extensive library of protein constructs to investigate the interactions of p53 and E1A with the TAZ1, TAZ2, KIX, and nuclear coactivator binding domains of CBP. The structures of the complexes formed by the bipartite N-terminal transactivation domain of p53 with the CBP domains will be determined, to obtain new insights into the dynamic interplay of interactions between the domains and into the molecular basis for CBP/p300 activation of p53-regulated transcription. The effects of phosphorylation of the p53 transactivation domain, at single and multiple sites, on interactions with these domains will be investigated. Structures will be determined for the complexes formed by E1A (both non-oncogenic AdV5 and strongly oncogenic AdV12 serotypes), with domains of CBP/p300, in order to obtain insights into the molecular basis by which E1A competes with other transcription factors to sequester CBP/p300 and subvert the cellular transcription machinery. Finally, two ternary complexes will be characterized, between HDM2, p53, and the TAZ1 domain of CBP, and between the pocket domain of the retinoblastoma protein Rb, E1A and the TAZ2 domain. With its emphasis on weak, competing interactions and the synergistic interplay between multiple binding sites and multiple partners, this research will provide new and fundamental insights into the mechanisms of molecular recognition by CBP/p300, and the molecular basis by which they mediate critical interactions with p53 and viral oncoproteins in tumor suppression, growth and progression.

描述（由申请人提供）：CREB结合蛋白（CBP）和p300是模块化转录共激活因子，在真核细胞中整合众多信号转导途径，并且对于生长、分化、凋亡和DNA修复至关重要。 CBP/p300 在胚胎发育中发挥着关键作用，并且是许多致癌事件的靶标。它们还起到肿瘤抑制因子的作用，并且是许多细胞和肿瘤病毒癌蛋白的转录活性所必需的。 CBP 和 p300 在介导肿瘤抑制因子 p53 的稳定性和转录活性方面发挥着重要作用。这些蛋白质之间的相互作用是高度动态的，并通过 p53 反式激活结构域在多个位点的磷酸化来调节，以响应基因毒性应激。腺病毒癌蛋白 E1A 与细胞因子竞争隔离 CBP/p300，破坏正常细胞周期控制以诱导细胞永生化。这项研究将利用广泛的蛋白质构建体库来研究 p53 和 E1A 与 CBP 的 TAZ1、TAZ2、KIX 和核共激活剂结合域的相互作用。将确定 p53 的二分 N 端反式激活结构域与 CBP 结构域形成的复合物的结构，以获得对结构域之间相互作用的动态相互作用以及 CBP/p300 激活 p53 调节转录的分子基础的新见解。将研究 p53 反式激活结构域在单个和多个位点的磷酸化对与这些结构域相互作用的影响。将确定 E1A（非致癌 AdV5 和强致癌 AdV12 血清型）与 CBP/p300 结构域形成的复合物的结构，以便深入了解 E1A 与其他转录因子竞争隔离 CBP/p300 并颠覆细胞转录机制的分子基础。最后，将表征两个三元复合物，即 HDM2、p53 和 CBP 的 TAZ1 结构域之间，以及视网膜母细胞瘤蛋白 Rb 的口袋结构域、E1A 和 TAZ2 结构域之间。这项研究强调弱的竞争性相互作用以及多个结合位点和多个伙伴之间的协同相互作用，将为 CBP/p300 的分子识别机制以及它们在肿瘤抑制、生长和进展中介导与 p53 和病毒癌蛋白关键相互作用的分子基础提供新的基本见解。