Structural basis for CBP/p300 transcriptional regulation

CBP/p300 转录调控的结构基础

基本信息

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
项目类别：
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

项目摘要

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，可以整合真核细胞中的许多信号转导途径，对于生长，分化，凋亡和DNA修复至关重要。 CBP/P300在胚胎发育中起关键作用，并且是许多致癌事件的目标。它们还充当肿瘤抑制子，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p53的稳定性和转录活性中起着至关重要的作用。这些蛋白质之间的相互作用是高度动态的，并且通过响应于遗传毒性应激的多个位点的p53反式激活结构域的磷酸化来调节。腺病毒癌蛋白E1A与细胞因子竞争，以隔离CBP/p300，破坏了正常的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与CBP结构域的两部分N末端反式化结构域形成的复合物的结构，以获取对域之间相互作用的动态相互作用的新见解，并以CBP/P300激活p53受调的转录的CBP/P300激活。将研究p53反式激活结构域的磷酸化，单位和多个位点与这些结构域相互作用的影响。将确定由E1A（非康复ADV5和强烈致癌adv12血清型）形成的复合物，具有CBP/P300的域，以获取E1A与其他转录因子竞争CBP/P300并删除细胞转录的分子基础的见解。最后，将在HDM2，p53和CBP的TAZ1域之间以及视网膜细胞瘤蛋白RB，E1A和TAZ2域之间表征两个三元复合物。 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.