A Novel Role for PML bodies in Leukemic Transformation

PML 体在白血病转化中的新作用

• 批准号: 8784600
• 负责人: Alexis A Cogswell
• 金额: $ 4.95万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8784600
• 关键词: Acute Promyelocytic Leukemia; Address; Affect; Aftercare; Apoptosis; Binding; Binding Proteins; Biological Assay; Cancerous; Cell Cycle Checkpoint; Cell Nucleolus; Cell Nucleus; Cell Proliferation; Cell physiology; Cells; Cellular Stress; Chimeric Proteins; DNA biosynthesis; Development; Disease; Gene Expression; Gene Proteins; Genome; Goals; Growth; Hela Cells; Immunoprecipitation; Interferons; Ionizing radiation; Lac Repressors; Link; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Membrane; Messenger RNA; Methods; Modeling; Molecular Biology; Molecular and Cellular Biology; Monitor; Normal Cell; Nuclear; Oncogene Proteins; Oncogenes; Organelles; Oxidative Stress; Peptide Elongation Factor 1; Play; Process; Protein Biosynthesis; Proteins; Quality Control; RARA gene; RNA Interference; Recruitment Activity; Regulation; Research; Retinoblastoma Protein; Ribosomal Proteins; Ribosomes; Role; Serum; Set protein; Site; Starvation; Stress; Structure; System; Testing; Transfection; Translating; Translations; Tumor Suppression; Tumor Suppressor Proteins; Work; base; biological adaptation to stress; cancer type; cell growth; cell transformation; design; insight; metaplastic cell transformation; novel; novel therapeutics; protein degradation; public health relevance; research study; response; senescence; therapeutic target; transcription factor PML

DESCRIPTION (provided by applicant): The central dogma of molecular biology describes the cellular processes that are involved in the replication of DNA and expression of genes within a genome. Once considered definitive, it has instead been continuously modified and refined, as new discoveries bring novel insights to our understanding of molecular and cellular biology. Recent studies show strong and convincing evidence that protein synthesis occurs in the nucleus; however, the functional significance of nuclear translation remains unknown. Unlike the compartmentalization of cytoplasmic functions into membrane-bound organelles, activities within the nucleus are contained in non- membranous assemblies, or nuclear bodies (NBs). Promyelocytic leukemia (PML) nuclear bodies have multiple proposed functions, but the precise role of PML bodies remains poorly characterized. Prior research suggests that PML bodies are involved in protein degradation, apoptosis, and the response to cellular stress. Additional evidence also indicates that PML bodies may play a role in protein synthesis. Several components of the translation machinery including ribosomal proteins, eIF3, eIF4E, and elongation factor 1 are found in PML bodies. Moreover, it is suggested that PML bodies require eIF4E for proper assembly. Based upon the overwhelming evidence that PML bodies contain translation machinery, we have tested and determined that there is indeed PML-mediated nuclear translation. This proposal will determine how PML-localized protein translation functions in the context of cellular stress and transformation, and will identify the proteins involved in translation at PML bodies. Aim 1 will determine how PML-mediated nuclear translation is altered by cell stress and transformation and investigate the link between nuclear translation and aberrant cell growth or proliferation. Aim 2 will identify the proteins present at translating PML bodies in normal, stressed, and transformed cells using immunoprecipitation and mass spectrometry. In tandem, the dynamic assembly of translating PML bodies will be interrogated to determine if proteins involved in translation can form PML bodies de novo. Aim 3 will determine if experimental manipulation of PML expression alters nuclear translation and the transformation efficiency thus establishing a novel role for PML and nuclear translation in disease. The experiments described in this proposal are designed to test the novel hypothesis that nuclear translation occurs at PML bodies and is involved in the regulation of the stress response and cellular transformation. This study also takes a unique approach to examine the mechanism of tumor suppression by PML through addressing the role of PML mediated nuclear translation in cell fate choices. Overall, these experiments will contribute to the understanding o a new process, nuclear translation, and may uncover previously unknown therapeutic targets for multiple cancers.

描述（由申请人提供）：分子生物学的中心法则描述了基因组中DNA复制和基因表达所涉及的细胞过程。它曾经被认为是确定的，但随着新的发现为我们对分子和细胞生物学的理解带来新的见解，它一直在不断地修改和完善。最近的研究显示了强有力的、令人信服的证据，证明蛋白质合成发生在细胞核中；然而，核翻译的功能意义尚不清楚。与细胞质功能被划分为膜结合细胞器不同，细胞核内的活动被包含在非膜组装或核体（NBs）中。早幼粒细胞白血病（PML）核小体具有多种功能，但PML小体的确切作用仍不清楚。先前的研究表明PML小体参与蛋白质降解、细胞凋亡和对细胞应激的反应。其他证据也表明PML小体可能在蛋白质合成中发挥作用。翻译机制的几个组成部分，包括核糖体蛋白、eIF3、eIF4E和延伸因子1，在PML小体中被发现。此外，还建议PML体需要eIF4E才能正确组装。基于PML体含有翻译机制的压倒性证据，我们已经测试并确定确实存在PML介导的核翻译。该提案将确定PML定位蛋白翻译如何在细胞应激和转化的背景下起作用，并将确定PML小体中参与翻译的蛋白质。目的1将确定pml介导的核翻译如何被细胞应激和转化所改变，并研究核翻译与异常细胞生长或增殖之间的联系。目的2将使用免疫沉淀和质谱法鉴定正常、应激和转化细胞中翻译PML小体的蛋白质。同时，翻译PML小体的动态组装将被询问，以确定参与翻译的蛋白质是否可以重新形成PML小体。目的3将确定PML表达的实验操作是否会改变核翻译和转化效率，从而建立PML和核翻译在疾病中的新作用。本实验旨在验证核翻译发生在PML小体中并参与应激反应和细胞转化的调节的新假设。本研究还采用了一种独特的方法，通过解决PML介导的核翻译在细胞命运选择中的作用来研究PML抑制肿瘤的机制。总的来说，这些实验将有助于理解一个新的过程，核翻译，并可能发现以前未知的多种癌症的治疗靶点。