The Structure and Function of Perinucleolar Compartment

核仁周围室的结构和功能

• 批准号: 8655542
• 负责人: Sui HUANG
• 金额: $ 30.9万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-10 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8655542
• 关键词: Binding; Boxing; Cancerous; Cartilage; Cell Cycle; Cell Differentiation process; Cell Nucleolus; Cell Nucleus; Cell physiology; Cells; ChIP-seq; Chromosomes; Complex; DNA; DNA Polymerase III; Data; Foundations; Functional RNA; Funding; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Glycerol; Goals; Grant; Hair; Label; Lead; Link; Malignant Neoplasms; Modeling; Molecular; Mutagenesis; Mutation; Neoplasm Metastasis; Nuclear Structure; Pathway interactions; Phenotype; Play; Process; Proteins; Protocols documentation; RNA; Regulation; Resolution; Ribonucleases; Role; Site; Structure; Testing; Work; base; cancer cell; cell assembly; cell growth; human disease; insight; interest; light microscopy; malignant phenotype; novel; protein complex; research study; yeast genetics

DESCRIPTION (provided by applicant): PNCs are structurally linked to the nucleoli, and they assemble and disassemble in concert with nucleoli through the cell cycle. It is highly enriched with newly synthesized pol III RNA and is dependent upon continuous pol III transcription. The PNC forms in metastatically capable cells and the assembly of the PNC is not simply the results of over expression of the PNC components. Over the last funding cycle (3.5 years), using RNase MRP RNA (RMRP) as an example, we have found that PNC contains a novel RMRP containing complex. The complex contains PTB, CUGBP, RMRP, and other unknown factors, and is large, pelleting at the bottom of a 5-40% glycerol gradient. Using a high resolution light microscopy (OMX), we found that RMRP-PTB-CUGB closely colocalize at the PNC. However, RMRP at the PNC does not colocalize with newly synthesized RNA, suggesting the RNP may not represent the post transcriptional processing of nascent RMRP. In addition, we found that the PNC is associated with unknown DNA loci. These findings lead to our current working model. Unknown RNPs become nucleated upon specific DNA loci, most likely a pol III gene(s), and regulates their expression in PNC containing cells. The regulation can be at the levels of DNA or RNA, or both. We propose three complementary aims to test this hypothesis and to investigate the structure and function of the PNC. We will identify the components of the novel RMRP-PTB-CUGBP complex and the functional relevance of this complex to the PNC and cellular activities. We will identify the PNC associated DNA loci and will dissect the relationship between the novel RNPs and the expression of the loci. The results of these experiments will not only provide the necessary foundation to the future understanding of the structure and function of the PNC and malignancy, but will also reveal uncharacterized function of highly conserved RMRP.

描述（由申请人提供）：pnc在结构上与核仁相连，它们在细胞周期中与核仁一起组装和拆卸。它富含新合成的pol III RNA，并依赖于连续的pol III转录。PNC在具有转移能力的细胞中形成，PNC的组装不仅仅是PNC成分过度表达的结果。在上一个资助周期（3.5年）中，以RNase MRP RNA （RMRP）为例，我们发现PNC含有一种新的含有RMRP的复合物。该复合物含有PTB、CUGBP、RMRP和其他未知因子，体积大，在5-40%甘油梯度的底部形成球团。利用高分辨率光学显微镜（OMX），我们发现RMRP-PTB-CUGB在PNC处紧密共定位。然而，PNC的RMRP不与新合成的RNA共定位，这表明RNP可能不代表新生RMRP的转录后加工。此外，我们发现PNC与未知的DNA位点相关。这些发现导致了我们目前的工作模型。未知的RNPs在特定的DNA位点（最有可能是pol III基因）上成核，并调节其在含PNC细胞中的表达。这种调节可以在DNA或RNA的水平上，或者两者都有。我们提出三个互补的目标来检验这一假设，并研究PNC的结构和功能。我们将鉴定新型RMRP-PTB-CUGBP复合物的成分以及该复合物与PNC和细胞活动的功能相关性。我们将鉴定与PNC相关的DNA位点，并剖析新型RNPs与该位点表达之间的关系。这些实验结果不仅将为进一步了解PNC和恶性肿瘤的结构和功能提供必要的基础，而且还将揭示高度保守的RMRP未被表征的功能。