EXPRESSION AND FUNCTION OF U1 RNA ISOFORMS DURING DEVELOPMENT

这一提议代表着我们继续努力， 有希望的少数民族学生寻求在生物医学科学的职业。 为了实现这一目标，学生们已经并将在所有培训 分子生物学方面。 这项科学计划 目的涉及U1的结构和功能表征- 家蚕的相关序列及其转录本。 U1 RNA是六种主要的核小RNA（snRNA）之一，U1-U6， 已知与6到10种蛋白质结合形成小的 核糖核蛋白颗粒（snRNP）参与转化 将hnRNA剪接成mRNA。 我们的实验室部分 在这种生物体中鉴定了三类U1相关基因。 一类是转录失活的假基因; 另一种，是由一个高度重复的U1相关序列所代表的， 发育和组织特异性转录，表明在 基因表达的调控。 本文中概述的实验 该提案将允许我们的学生阐明U1亚型发挥的作用 在RNA剪接水平上调节基因表达。 他们的 参与hnRNA剪接将使用发育 分阶段的丝腺、滤泡和几种细胞系。 U1亚型将 纯化并在体外用于重构实验，其中 它们差异组装成snRNP/hnRNP颗粒，与 含有内含子的丝心蛋白和绒毛膜RNA，剪接能力将 追究 将进行体内实验以研究 U1亚型在这些细胞中的差异组装和剪接 分阶段的细胞类型和细胞系。 与β地中海贫血的情况一样， 人类中的RNA剪接突变， U1亚型的效率和/或特异性可能被证明是 一些人类疾病 蚕蛾，具有良好的学习发育能力 分子生物学，加上高度专业化的细胞类型， 独特的机会，让我们的学生检查这些U1的功能- 相关序列 具体而言，少数民族学生将被激励， 通过让他们接触生物医学科学， 相关文献，并训练他们设计实验，制定 协议，进行实验并公布结果。

This proposal represents a continuation of our efforts to motivate promising minority students to seek careers in the biomedical sciences. To achieve this goal, students have been and will be trained in all aspects of molecular biology. The scientific project employed for this purpose involves the structural and functional characterization of U1- related sequences and their transcripts in the silkmoth Bombyx mori. U1 RNA is one of six prominent small nuclear RNAs (snRNAs), U1-U6, which are known to associate with six to ten proteins to form small ribonucleoprotein particles (snRNPs) which participate in the conversion of hnRNA to mRNA by splicing. Our laboratory has partially characterized three categories of U1-related genes in this organism. One category is composed of transcriptionally inactive pseudogenes; another, is represented by a highly reiterated U1-related sequences are developmentally and tissue-specific transcribed, suggesting a role in the regulation of gene expression. The experiments outlined in this proposal will allow our students to elucidate the role U1 isoforms play in the regulation of gene expression at the RNA splicing level. Their involvement in hnRNA splicing will be determined using developmentally staged silk glands, follicles, and several cell lines. U1 isoforms will be purified and used in vitro in reconstitution experiments in which their differential assembly into snRNP/hnRNP particles, association with intron-containing fibroin and chorion RNAs, the splicing capacities will be investigated. In vivo experiments will be performed to study the differential assembly and splicing efficiently of U1 isoforms in these staged cell types and cell lines. As with the case of beta-thalassemia RNA splicing mutations in humans, differential splicing as a result of U1 isoforms efficiency and/or specificity may prove to be the basis of some human maladies. The silkmoth , with a well-studies developmental and molecular biology, plus highly specialized cell types, provides a unique opportunity for our students to examine the function of these U1- related sequences. Specifically, minority students will be motivated to pursue a career in the biomedical sciences by exposing them to the pertinent literature, and training them to design experiments, formulate protocols, conduct experiments and publish the results.