Analysis of microtubule localized RNAs

6.摘要 该提案的长期目标是确定靶向的顺式和反式作用系统。 有丝分裂微管的特异性RNA，并了解有丝分裂过程中局部RNA的功能 发展先行者的要求在细胞分裂期间遗传物质的稳定遗传对于细胞的生长至关重要。 所有生物体的生存和分化。RNA在细胞内具有多种功能， 既作为mRNA形式的遗传信息，又作为各种基因的结构和催化成分， 蛋白质复合物mRNA和非编码RNA的转录后调节 转运到特定的亚细胞位置是调节基因表达的广泛使用的机制 在时间和空间上。不同类型的RNA在有丝分裂纺锤体上的定位是一个很好的试验 了解本地化的mRNA如何控制有丝分裂纺锤体功能和遗传的情况 遗传物质该提案将使用体外和体内试验的组合来鉴定 在减数分裂期间将特异性mRNA靶向有丝分裂纺锤体的顺式作用定位序列， 在整个发展中。这些信息将大大增加已知的 本地化元素，并提供所需的信息，以建立准确的模型， 定位RNA的预测。关于mRNA定位元件的信息将用于 生物化学纯化和鉴定负责RNA转运的反式作用蛋白因子， 微管，增加了对用于运输特定RNA的系统类型的理解， 细胞或有机体中的离散位置。最后，定位于有丝分裂细胞的mRNA的特性 纺锤体表明定位翻译可能在各种有丝分裂事件调节中起主要作用。 这项建议将研究有丝分裂翻译在细胞周期调控中的作用，这将提供 深入了解细胞周期的翻译控制。这将是第一个全面的 对定位于特定亚细胞结构的mRNA的分析，将提供对系统的深入了解 用于mRNA定位，并提供有关定位的mRNA的功能作用， 细胞周期调控这里描述的实验方法将普遍适用于 了解局部RNA在各种细胞过程中的影响。7.

6. Abstract 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.