Analysis of microtubule localized RNAs

描述(由申请人提供)：这项建议的长期目标是确定针对特定RNA到有丝分裂微管的顺式和反式作用系统，并了解局部RNA在有丝分裂和发育过程中的功能。细胞分裂过程中遗传物质的稳定遗传对所有有机体的生存和分化至关重要。RNA在细胞内具有广泛的功能，既是以mRNA形式存在的遗传信息，也是各种蛋白质复合体的结构和催化成分。通过运输到特定的亚细胞位置来对mRNAs和非编码RNAs进行转录后调控是一种广泛使用的调控基因表达的时间和空间机制。不同类型的RNA对有丝分裂纺锤体的定位是理解定位的mRNAs如何控制有丝分裂纺锤体功能和遗传物质遗传的一个很好的测试案例。这项建议将使用体外和体内试验相结合的方法来鉴定在减数分裂和整个发育过程中针对有丝分裂纺锤体的特定mRNAs的顺式作用定位序列。这些信息将大大增加已知的定位元件的数量，并提供为定位RNA的从头预测建立准确模型所需的信息。有关信使核糖核酸定位元件的信息将被用于生化提纯和鉴定负责将核糖核酸运输到微管的反式作用蛋白因子，从而增加对用于将特定核糖核酸运输到细胞或生物体内特定位置的系统类型的了解。最后，定位于有丝分裂纺锤体的mRNAs表明，局部翻译可能在各种有丝分裂事件的调控中发挥重要作用。这项建议将研究有丝分裂翻译在细胞周期调节中的作用，这将为深入了解细胞周期的翻译控制提供洞察力。这一建议将提供第一次对定位于特定亚细胞结构的mRNAs的全面分析，将提供对用于mRNAs定位的系统的洞察，并提供关于定位的mRNAs在细胞周期控制中的功能作用的信息。这里描述的实验方法将通常适用于理解定位的RNA在各种细胞过程中的影响。7.与公共卫生的相关性：这个项目的目标是了解mRNAs的定位如何影响细胞分裂过程中的微管组装和染色体分离。这项研究可能会让我们了解细胞分裂的机制，这些机制与癌症和唐氏综合症等人类疾病密切相关。更多地了解细胞分裂的机制可能会导致改进药物设计，从而影响人类健康。

DESCRIPTION (provided by applicant): The long-term goal of this proposal is to identify the cis and trans acting systems that target specific RNAs to mitotic microtubules and to understand the function of localized RNAs during mitosis and development. The stable inheritance of genetic material during cell division is critical for the survival and differentiation of all organisims. RNA serves a wide variety of functions within cells, acting both as genetic information in the form of mRNA, and as a structural and catalytic component of various protein complexes. Posttranscriptional regulation of both mRNAs and noncoding RNAs through transport to specific subcellular locations is a widely used mechanism for modulating gene expression in time and space. The localization of different types of RNA to the mitotic spindle is an excellent test case for understanding how localized mRNAs control mitotic spindle function and the inheritance of genetic material. This proposal will use a combination of in vitro and in vivo assays to identify both the cis acting localization sequences that target specific mRNAs to the mitotic spindle during meiosis and throughout development. This information will provide a large increase in the number of known localization elements and provide the information required to build accurate models for the de novo prediction of localized RNAs. Information about mRNA localization elements will be used to biochemically purify and identify trans acting protein factors responsible for the transport of RNA to microtubules, increasing the understanding of the types of systems used to transport specific RNAs to discreet locations within a cell or organism. Finally, the identities of mRNAs localized to the mitotic spindle suggest that localized translation might play a major role in regulation of various mitotic events. This proposal will examine the role of mitotic translation in cell cycle regulation, which will provide insight into translational control of the cell cycle. This proposal will provide the first comprehensive analysis of mRNAs localized to a specific subcellular structure, will provide insight into the systems utilized for mRNA localization, and provide information about the functional role of localized mRNAs in cell cycle control. The experimental approach described here will be generally applicable to understanding the impact of localized RNAs in various cellular processes. 7. Public Health Relevance: The goal of this project is to understand how localization of mRNAs affects microtubule assembly and chromosome segregation during cell division. This study is likely to inform our understanding of the mechanisms of cell division, which are intimately linked to human diseases such as cancer and Down's syndrome. Greater understanding of the mechanisms cell division may lead to improved drug design that could impact human health.