ORGANIZATION OF RIBONUCLEOPROTEINS IN MAMMALIAN CELLS

描述：(改编自申请人的摘要)：总体目标 调查员的实验室的中心是如何将 哺乳动物核影响基因表达的调控。核糖核酸 加工反应是基因途径中至关重要的一环 表达，以及小核核糖核蛋白(SnRNPs) 显示在RNA加工反应中起关键作用。核糖核蛋白 是风湿性疾病自身免疫反应的常见靶点 如红斑狼疮、硬皮病和干燥综合征。自.以来 了解基因和基因的组织和核“邮编” 他们在体内的产品需要能够将整个 细胞，他们选择使用荧光原位杂交(FISH) 和数字成像显微镜研究细胞内分布 核糖核蛋白(RNPs)和染色质(DNA)。与 在分析RNA加工机制方面取得的进展 在体外，人们对这些因素是如何作用的知之甚少。 在体内组织，或加工如何与转录整合， 聚腺苷酸化和转运。RNPs在细胞生物学中扮演着重要的角色。 通过定义这些复合体的相对细胞内位置 其他亚细胞组件或地标，研究人员的 对RNP参与的细胞过程的洞察。 该提案的两个具体目标是：(1)识别染色体 在哺乳动物细胞核中与卷曲小体共存的基因座 了解这些关联的性质。螺旋体(CBS)是 核细胞器的功能未知，其中的元素(主要是RNPs) 三条主要的RNA加工途径：前信使核糖核酸、组蛋白 MRNA和rRNA的加工。此外，他们还发现哥伦比亚广播公司 经常与编码组蛋白的基因簇相关， 间期HeLa细胞中的U1和U2 RNA。建议进行实验，以 确定这些关联的性质。(2)刻画 前驱病毒的结构组成和分子生理学 未确认的核亚区。两类小聚合酶III RNAs，包括Ro RNAs，集中在这种新的结构中 称之为核周室(PNC)。作为这一目标的一部分，他将 确定Ro RNPs的亚细胞位置，确定其他成分 它可能在PNC中共存，并调查RNP 在PNC内可穿梭于胞核和胞浆之间。 调查员在及时性方面处于突出地位 和专业知识来研究这些RNPs的亚细胞组织 令人着迷的核亚区并将它们的分布与 具有特定染色体序列的基因。总而言之，这些研究应该 对职能组织有新的、有趣的见解 哺乳动物的细胞核。

DESCRIPTION: (Adapted from the applicant's abstract): The overall goal of the investigator's laboratory centers on how the architecture of the mammalian nucleus influences the control of gene expression. RNA processing reactions form a vital link in the pathway of gene expression, and small nuclear ribonucleoproteins (snRNPs) have been shown to play key roles in RNA processing reactions. Ribonucleoproteins are frequent targets for autoimmune responses in rheumatic disorders such as lupus erythematosus, scleroderma and Sjogren's syndrome. Since understanding the organization and nuclear "zip-coding" of genes and their products in vivo requires techniques which can visualize whole cells, they have chosen to use fluorescence in situ hybridization (FISH) and digital imaging microscopy to study the intracellular distribution of ribonucleoproteins (RNPs) and chromatin (DNA). In contrast with the progress that has been made in analyzing the RNA processing machinery in vitro, relatively little is known about how these factors are organized in vivo or how processing is integrated with transcription, poladenylation and transport. RNPs play vital roles in cell biology. By defining the intracellular locations of these complexes relative to other subcellular components or landmarks, the investigator's will gain insights into the cellular processes in which RNPs participate. The two specific aims of the proposal are: (1) To identify chromosomal loci that colocalize with coiled bodies in mammalian cell nuclei and to understand the nature of these associations. Coiled bodies (CBs) are nuclear organelles of unknown function in which elements (mainly RNPs) of three major RNA processing pathways accumulate: pre-mRNA, histone mRNA and rRNA processing. In addition, they have found that CBs are frequently associated with the gene clusters that encode the histone, U1 and U2 RNAs in interphase HeLa cells. Experiments are proposed to identify the nature of these associations. (2) To characterize the structural composition and molecular physiology of a previously unidentified nuclear subdomain. Two classes of small polymerase III RNAs, including the Ro RNAs, concentrate within this novel structure we call the perinucleolar compartment (PNC). As part of this aim, he will determine the subcellular location of Ro RNPs, identify other components which may colocalize in PNCs and investigate the possibility that RNPs within the PNC may shuttle between the nucleus and the cytoplasm. The investigator's are in an outstanding position in terms of timeliness and expertise to study the subcellular organization of RNPs within these fascinating nuclear subdomains and to correlate their distributions with those of specific chromosomal sequences. Together, these studies should lead to new and interesting insights into the functional organization of the mammalian nucleus.