Analysis of microtubule localized RNAs

描述（由申请人提供）：本提案的长期目标是鉴定将特异性RNA靶向有丝分裂微管的顺式和反式作用系统，并了解有丝分裂和发育过程中局部RNA的功能。细胞分裂过程中遗传物质的稳定遗传对所有生物体的生存和分化至关重要。RNA在细胞内具有多种功能，既作为mRNA形式的遗传信息，又作为各种蛋白质复合物的结构和催化组分。mRNA和非编码RNA通过转运到特定的亚细胞位置进行转录后调节是一种广泛使用的在时间和空间上调节基因表达的机制。不同类型的RNA定位于有丝分裂纺锤体是了解定位的mRNA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。该提案将使用体外和体内测定的组合来鉴定在减数分裂期间和整个发育期间将特异性mRNA靶向有丝分裂纺锤体的顺式作用定位序列。这些信息将大大增加已知定位元件的数量，并提供建立精确模型所需的信息，用于从头预测定位的RNA。有关mRNA定位元件的信息将用于生物化学纯化和鉴定负责将RNA转运到微管的反式作用蛋白因子，增加对用于将特定RNA转运到细胞或生物体内离散位置的系统类型的理解。最后，定位于有丝分裂纺锤体的mRNA的身份表明，本地化的翻译可能在各种有丝分裂事件的调节中发挥重要作用。该提案将研究有丝分裂翻译在细胞周期调控中的作用，这将为细胞周期的翻译控制提供见解。该提案将提供定位于特定亚细胞结构的mRNA的第一次全面分析，将提供用于mRNA定位的系统的洞察力，并提供有关定位的mRNA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普遍适用于理解本地化RNA在各种细胞过程中的影响。7.公共卫生相关性：本项目的目标是了解mRNA的定位如何影响微管组装和细胞分裂过程中的染色体分离。这项研究可能会让我们了解细胞分裂的机制，这与癌症和唐氏综合症等人类疾病密切相关。更好地理解细胞分裂的机制可能会导致改善药物设计，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.