The regulation of PP1 in the nucleus

细胞核中PP1的调节

• 批准号: 9104450
• 负责人: Rebecca Page
• 金额: $ 11.44万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9104450
• 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; Up-Regulation; Work; X-Ray Crystallography; Yeasts; aurora B kinase; base; chromatin remodeling; daughter cell; fitness; genetic regulatory protein; human disease; insight; member; novel; premature; prevent; protein complex; public health relevance; research study; stem; structural biology

 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（由适用提供）：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适应性至关重要。相反，与有丝分裂相关的任何步骤（在父母细胞分为两个相同的子细胞的过程中）的破坏通常与疾病，尤其是癌症相关。快速新兴的情况是有丝分裂是由细胞周期激酶和磷酸酶（尤其是SER/THR磷酸酶蛋白磷酸酶1（PP1）之间的复杂平衡驱动的。尽管PP1本身具有广泛的特异性，但它通过形成稳定的复合物（Holoenzymes），其具有指导其活性和本地化的稳定复合物（Holoenzymes）的作用。我们的漫长目标是了解控制健康和疾病中PP1活性的结构和功能机制，这是我们实验室做出许多基本贡献的领域。在这里，我们描述了一个完整的研究计划，以了解核us中PP1的调节。提出的研究项目采用了强大的综合方法，该方法将X射线晶体学和NMR光谱与生化和细胞生物学实验结合在一起，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。 Specifically, we are focusing on the regulation of PP1 by four PP1-targeting proteins: 1) Repoman (recruits PP1 onto mitotic chromatin at ananaphase), 2) PNUTS (PP1 nuclear targeting subunit), 3) Knl1 (Kinetochore null protein 1) and 4) the ASPP (Apoptosis-stimulating of p53 protein) family of proteins.这些调节剂中每个调节剂的异常表达与多种癌症的发病率和严重程度增加有关。这些调节剂与当前缺少的PP1分子相互作用的详细描述是对这些重要全酶的全面理解。在我们的综合努力中，我们将：1）确定这些PP1调节剂的自由形式的结构，2）确定PP1全酶的结构，3）确定这些复合物如何直接和调节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这些关键核PP1全酶的生物学函数和作用模式。该提案中描述的研究利用了PP1研究领域领先的研究人员的广泛专家，并利用了最好的国家和国际合作者。最后，它的初步数据表明，这项工作将为PP1的分子调节及其在细胞周期的不同阶段提供独特的新颖见解。