The Structure and Function of Perinucleolar Compartment

描述（由申请人提供）：我们的长期目标是了解PNC的结构和功能及其在恶性肿瘤中的作用。核周室（PNC）是一种多组分的非膜结合核亚结构，位于核仁周围。我们发现它在体内和体外的恶性转化中都是独一无二的。PNC富含由pol III新合成的RNA和主要参与pol II RNA加工的RNA结合蛋白。我们的初步研究结果表明，pnc相关RNA与一些已知的pol II RNA结合蛋白处于相同的复合物中。这些新的复合物并不局限于PNC，因为PTB-MRP RNA相互作用也可以在没有PNC的正常细胞中检测到。然而，这些复合物的水平或调控可能在恶性肿瘤期间发生变化，导致这些复合物在PNC中成核。这些发现导致了我们的工作模型，其中PNC与恶性肿瘤期间新合成的pol III RNA代谢相关的新分子复合物的变化有关，并且可能代表了这种变化。本提案的目标是通过识别和初步表征这些新的RNPs来开始测试这一假设，使用RNase MRP RNA （PNC的真正成分之一）作为第一个例子，并开始使用细胞生物学方法解决这些RNPs与PNC的功能关联。由于人们对pol III小非编码rna的转录后加工和调控知之甚少，因此新rna的解决不仅对理解PNC的结构和功能很重要，而且还应该揭示这些功能rna的转录后调控的新机制。

DESCRIPTION (provided by applicant): Our long-term goal is to understand the structure and function of the PNC and its roles in malignancy. The perinucleolar compartment (PNC) is a multicomponent non-membrane bound nuclear substructure that localizes to the nucleolar periphery. We have found it to be unique to malignant transformation both in vitro and in vivo. The PNC is enriched with RNA newly synthesized by pol III and RNA binding proteins primarily implicated in pol II RNA processing. Our preliminary findings show that the PNC-associated RNAs are in the same complexes with some of the known pol II RNA binding proteins. These novel complexes are not restricted to the PNC since PTB-MRP RNA interactions can also be detected in normal cells without PNCs. However, the levels or the regulations of these complexes may change during malignancy, leading to the nucleation of these complexes in the PNC. These findings lead to our working model in which the PNC is associated with and may represent changes in novel molecular complexes that are involved in the metabolism of newly synthesized pol III RNA during malignancy. The goal of this proposal is to begin test this hypothesis by identifying and initially characterizing these novel RNPs, using RNase MRP RNA (one of the bona fide components of the PNC) as the first example, and by beginning to address the functional association of these RNPs with the PNC using cell biological approaches. As little is understood regarding the post-transcriptional processing and regulation of the pol III small non-coding RNAs, the solution of the novel RNPs is not only important for the understanding of PNC structure and function, but should also reveal a novel mechanism involved in the post-transcriptional regulation of these functional RNAs.