Regulation of ARF tumor suppressor function by biological phase separation

通过生物相分离调节ARF肿瘤抑制功能

基本信息

批准号：
10065965
负责人：
Eric B Gibbs
金额：
$ 6.74万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-09 至 2021-07-08
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10065965
关键词：
Affect Amino Acid Sequence Apoptosis Arginine Behavior Binding Biogenesis Biological Assay Biological Models Biological Process Biomolecular Nuclear Magnetic Resonance CDKN2A gene Cancer Cell Growth Cell Count Cell Cycle Arrest Cell Nucleolus Cells Cellular Stress Cellular biology Characteristics Complex Event Exhibits Fluorescence Fluorescence Recovery After Photobleaching Future Growth Homeostasis Human Hydrophobicity Image Imaging Techniques In Vitro Interphase Length Link Liquid substance MDM2 gene Maintenance Malignant - descriptor Malignant Neoplasms Maps Mediating Methods Microscopy Modeling Molecular Monitor Mouse Protein Mus NMR Spectroscopy NPM1 gene Neutrons Normal Cell Nuclear Export Nucleolar Proteins Oncogenes Oncogenic Phase Phenotype Play Production Property Protein Biosynthesis Proteins RNA Reading Regulation Ribosomal RNA Ribosomes Site Site-Directed Mutagenesis Solid Spectrum Analysis Stress Structure System TP53 gene Techniques Testing Time Training Tumor Suppressor Genes Tumor Suppressor Proteins biological adaptation to stress biophysical model cancer cell cell growth experience experimental study fluorescence imaging frontier insight live cell imaging loss of function molecular modeling mutant novel novel therapeutic intervention nucleophosmin p19ARF particle polysome profiling prevent reconstitution sensor solid state nuclear magnetic resonance structural biology targeted cancer therapy tumor

项目摘要

PROJECT SUMMARY The proposed study aims to determine how phase separation regulates the ARF protein, a tumor suppressor that frequently experiences loss of function in human cancers. During oncogenic stress, ARF sequesters essential proteins in the nucleolus in order to arrest the cell cycle or induce apoptosis. Specifically, ARF sequesters HDM2 in the nucleolus, which activates p53-dependent cell cycle arrest or apoptosis. ARF also sequesters NPM1 in the nucleolus, disrupting ribosome assembly, which also induces cell cycle arrest independently of p53. However, the mechanism by which ARF sequesters its targets in the nucleolus is not understood. Recent observations have demonstrated that nucleoli possess liquid-like properties, which manifest through phase separation of their constituent proteins and RNAs. Recently, our lab along with the Brangwynne group, showed that the liquid like features of the nucleolus depends in-part on the phase separation properties of NPM1. I hypothesize that p14ARF disrupts the liquid-like properties of the granular component through hydrophobic self-association and multivalent interactions of its multiple arginine-rich motifs with NPM1, resulting in inhibition of ribosome biogenesis. This study will determine (1) how p14ARF’s physicochemical properties allow it to sequester its target proteins within phase separated bodies, (2) the structure and dynamics of phase separated p14ARF-NPM1 complexes, and (3) the effect of p14ARF expression on NPM1 dynamics, ribosome biogenesis and growth in live cells. Results from this study will provide novel insights into ARF’s nucleolar tumor suppressor function, and in a broader context, how the fluid-features of nucleoli are altered during oncogenic stress events.

项目摘要拟议的研究旨在确定相位分离如何调节ARF蛋白，肿瘤抑制剂这种经常会在人类癌症中丧失功能。在致癌应力期间，ARF隔离器核仁中必需的蛋白质是为了阻止细胞周期或诱导凋亡。具体而言，ARF Nucleolus中的HDM2隔离，该HDM2激活p53依赖性细胞周期停滞或凋亡。也是ARF 隔离核糖中的NPM1，破坏了核糖体组装，这也诱导细胞周期停滞独立于p53。但是，ARF隔离其在核仁中的机制不是理解。最近的观察结果表明，核仁具有液体样性能，通过其组成蛋白和RNA的相位分离。最近，我们的实验室与Brangwynne集团一起结果表明，核仁的液体类似特征在部分取决于NPM1的相分离特性。我假设p14arf通过疏水性自缔合和其多种精氨酸的基序与NPM1的多价相互作用，导致核糖体生物发生抑制。这项研究将确定（1）P14ARF的物理特性如何隔离其目标相位分离物体内的蛋白质，（2）相分离的P14ARF-NPM1的结构和动力学复合物和（3）P14ARF表达对NPM1动力学，核糖体生物发生和生长的影响在活细胞中。这项研究的结果将为ARF的核肿瘤抑制剂功能提供新的见解，在更广泛的背景下，在致癌应力事件中如何改变核的流体功能。