The Structure and Function of Perinucleolar Compartment

核仁周围室的结构和功能

• 批准号: 8451354
• 负责人: Sui HUANG
• 金额: $ 29.82万
• 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/8451354
• 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，并且它们在整个细胞周期中与核仁一起组装和分解。它富含新合成的 pol III RNA，并且依赖于连续的 pol III 转录。 PNC 在具有转移能力的细胞中形成，并且 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含有一种新型的RMRP复合物。该复合物含有 PTB、CUGBP、RMRP 和其他未知因子，并且体积较大，在 5-40% 甘油梯度的底部沉淀。使用高分辨率光学显微镜 (OMX)，我们发现 RMRP-PTB-CUGB 紧密共定位于 PNC。然而，PNC 处的 RMRP 并不与新合成的 RNA 共定位，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当前的工作模式。未知的 RNP 在特定 DNA 位点（最有可能是 pol III 基因）上成核，并调节它们在含有 PNC 的细胞中的表达。调节可以在 DNA 或 RNA 水平上进行，或者在两者水平上进行。我们提出了三个互补的目标来检验这一假设并研究 PNC 的结构和功能。我们将确定新型 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 PNC 相关 DNA 基因座，并剖析新的 RNP 与基因座表达之间的关系。这些实验结果不仅将为未来理解PNC的结构和功能以及恶性肿瘤提供必要的基础，而且还将揭示高度保守的RMRP的未表征的功能。