The regulation of PP1 in the nucleus

细胞核中PP1的调节

基本信息

批准号：
9323496
负责人：
Rebecca Page
金额：
$ 32.24万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-11 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9323496
关键词：
Amino Acids Anaphase Antineoplastic Agents Apoptosis Area Behavior Binding Binding Proteins Biochemical Biochemistry and Cellular Biology Biological Process C-terminal Cell Cycle Cell Cycle Stage Cell Nucleus Cell division Cells Cellular biology Chromatin Chromosomes Complex Conflict (Psychology)DNA DNA Binding Domain Data Disease Down-Regulation Drug Targeting Equilibrium Eukaryota Event Genetic Transcription Genome Genomics Goals HMGB Proteins Health Holoenzymes Incidence International Kinetochores Laboratories Malignant Neoplasms Mediating Metaphase Mitosis Mitotic Molecular N-terminal NMR Spectroscopy Nuclear Nuclear Protein Phosphatase Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Phosphotransferases Process Proline-Rich Domain Protein Dephosphorylation Protein Family Protein p53 Protein phosphatase Proteins Reaction Recruitment Activity Regulation Reporting Research Research Personnel Research Project Grants Role SH3 Domains Severities Signal Pathway Site Specificity Structure TP53 gene Time Up-Regulation Work X-Ray Crystallography Yeasts aurora B kinase chromatin remodeling daughter cell experimental study fitness genetic regulatory protein human disease insight member novel premature prevent protein complex protein structure public health relevance stem structural biology

项目摘要

DESCRIPTION (provided by applicant): The accurate distribution of the replicated genome during cell division is crucial for genomic inheritance and cellular fitness. Conversely, disruptios in any of the steps associated with mitosis (the process in which a parental cell divides into two identical daughter cells) are often correlated with disease, especially cancer. The rapidly emerging picture is that mitosis is driven by the intricate balance between cell cycle kinases and phosphatases, especially the ser/thr phosphatase protein phosphatase 1 (PP1). While PP1 itself has a broad specificity, it acts in a highly specific manner by forming stable complexes (holoenzymes) with a host of regulatory proteins that direct its activity and localization. Our lon-term goal is to understand the structural and functional mechanisms that control PP1 activity in health and disease, an area in which our laboratory has made many fundamental contributions. Here we describe a complete research plan to understand the regulation of PP1 in the nucleus. The presented research project uses a powerful integrated approach that combines X-ray crystallography and NMR spectroscopy with biochemical and cell biology experiments to obtain novel insights into the molecular mechanisms that regulate PP1 activity during distinct stages of the cell cycle. Specifically, we are focusing on the regulation of PP1 by four PP1-targeting proteins: 1) Repoman (recruits PP1 onto mitotic chromatin at anaphase), 2) PNUTS (PP1 nuclear targeting subunit), 3) Knl1 (Kinetochore null protein 1) and 4) the ASPP (Apoptosis-stimulating of p53 protein) family of proteins. Abnormal expression of each of these regulators is associated with the increased incidence and severity of multiple cancers. Detailed descriptions of the molecular interactions of these regulators with PP1, which are currently missing, are needed for a comprehensive functional understanding of these important holoenzymes. In our combined efforts, we will: 1) determine the structures of the free form of these PP1 regulators, 2) determine the structures of the PP1 holoenzymes and 3) determine how these complexes direct and regulate PP1 activity. We will then leverage these protein and protein complex structures to elucidate, at a molecular level, the biological functions and modes of action of these key nuclear PP1 holoenzymes. The research described in this proposal leverages the extensive expertise of leading investigators in the PP1 research field, as well as takes advantage of the best possible national and international collaborators. Finally, it has the preliminary data that demonstrates that this work will provide unique, novel insights into the molecular regulation of PP1 and its fundamental roles during distinct stages of the cell cycle.

描述（由申请人提供）：细胞分裂过程中复制基因组的准确分布对于基因组遗传和细胞离线适应性至关重要。与有丝分裂（亲代细胞分裂成两个相同子代细胞的过程）相关的任何步骤的破坏细胞）通常与疾病，特别是癌症相关，迅速出现的情况是有丝分裂是由细胞周期激酶和磷酸酶（尤其是丝氨酸/苏氨酸磷酸酶蛋白）之间的复杂平衡驱动的。虽然 PP1 本身具有广泛的特异性，但它通过与指导其活性和定位的大量调节蛋白形成稳定的复合物（全酶）来以高度特异性的方式发挥作用。控制健康和疾病中 PP1 活性的结构和功能机制，我们的实验室在这一领域做出了许多基础性贡献。在这里，我们描述了一个完整的研究计划，以了解 PP1 在细胞核中的调节。一种强大的综合方法，将 X 射线晶体学和核磁共振波谱与生化和细胞生物学实验相结合，以获得对细胞周期不同阶段调节 PP1 活性的分子机制的新见解。四种 PP1 靶向蛋白：1) Repoman（将 PP1 招募到有丝分裂后期染色质上）、2) PNUTS（PP1 核靶向亚基）、3) Knl1 （着丝粒无效蛋白 1）和 4）ASPP（p53 蛋白凋亡刺激蛋白）家族蛋白的异常表达与多种癌症的发病率和严重程度增加有关。为了全面了解这些重要的全酶的功能，需要使用目前缺失的 PP1 调节因子。在我们的共同努力下，我们将：1) 确定这些游离形式的结构。 PP1 调节剂，2) 确定 PP1 全酶的结构，3) 确定这些复合物如何指导和调节 PP1 活性，然后我们将利用这些蛋白质和蛋白质复合物结构在分子水平上阐明生物学功能和作用模式。本提案中描述的研究利用了 PP1 研究领域领先研究人员的广泛专业知识，并利用了最好的国内和国际合作者。最后，初步数据表明这项工作将为 PP1 的分子调控及其在细胞周期不同阶段的基本作用提供独特、新颖的见解。