REGULATION OF PRE-MRNA SPLICING

mRNA 前剪接的调控

• 批准号: 6575602
• 负责人: Adrian R Krainer
• 金额: $ 22.84万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-21 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6575602
• 关键词: DNA binding protein; DNA footprinting; HeLa cells; RNA binding protein; RNA splicing; antisense nucleic acid; biological signal transduction; gel mobility shift assay; gene mutation; genetic library; genetic regulation; genetic screening; heterogeneous nuclear ribonucleoprotein; human immunodeficiency virus 1; immunoprecipitation; laboratory mouse; laboratory rabbit; molecular oncology; monoclonal antibody; neoplasm /cancer genetics; oncogenes; phosphorylation; precursor mRNA; protein localization; protein structure function; telomere; virus genetics

DESCRIPTION (provided by applicant): The control of gene expression at the post-transcriptional level is a fundamental problem in biology, with relevance to cancer. The basic mechanisms for the regulation of alternative splicing of cellular and viral genes in different tissues, developmental stages, or in response to external signals, will be investigated. We will carry out detailed structural and functional studies of hnRNP A1 and its derivatives, focusing on its nucleic acid-binding properties' its activities in alternative 5' splice-site selection, splicing silencing and telomere-length regulation, and the regulation of its localization and activity in response to genotoxic stress. We will carry out genetic screens in cultured mammalian cells to identify protein components involved in the fidelity of pre-mRNA splicing. Finally, we will continue the development of new technology involving rational design of compounds that mimic one of the activities of SR proteins, the activation of splicing via exonic splicing enhancers. These compounds will be tested for their ability to correct specific splicing defects caused by mutations in cancer susceptibility genes, and to modulate the expression of alternatively spliced isoforms of an apoptosis gene. These studies are directly relevant to the overall goals of the Program Project. As global regulators of alternative pre-mRNA splicing, hnRNP A1 and its antagonists, the SR proteins, may be responsible for the observed aberrant patterns of mRNA expression of numerous genes in transformed cells. Among potential targets of these regulators are several critical genes involved in the establishment or maintenance of the transformed phenotype, or in progression of malignancy. Regulation of alternative splicing is responsible for generating oncongenic and non-oncogenic forms of many cellular and viral oncogenes. Therefore, a better understanding of the basic mechanisms of alternative splicing regulation, and of the fidelity of this process, may lead to the design of drugs to specifically affect the synthesis of particular protein isoforms that play critical roles in tumorigenesis.

描述（由申请人提供）：基因表达的控制 转录后水平是生物学中的一个基本问题，具有相关性 到癌症。选择性剪接调控的基本机制 不同组织、发育阶段或不同阶段的细胞和病毒基因 对外部信号的响应，将被研究。我们将进行详细的 hnRNP A1 及其衍生物的结构和功能研究，重点是 它的核酸结合特性' 它在替代5'中的活性 剪接位点选择、剪接沉默和端粒长度调节，以及 其定位和活性的调节以响应遗传毒性 压力。我们将在培养的哺乳动物细胞中进行遗传筛选 鉴定参与前体 mRNA 剪接保真度的蛋白质成分。 最后，我们将继续开发涉及理性的新技术 设计模拟 SR 蛋白活性之一的化合物， 通过外显子剪接增强子激活剪接。这些化合物将 测试了其纠正由以下原因引起的特定拼接缺陷的能力 癌症易感基因突变，并调节其表达 凋亡基因的选择性剪接异构体。这些研究是 与计划项目的总体目标直接相关。 作为选择性前 mRNA 剪接的全局调节因子，hnRNP A1 及其 拮抗剂 SR 蛋白可能是观察到的异常现象的原因 转化细胞中许多基因的 mRNA 表达模式。之中 这些调节因子的潜在目标是参与的几个关键基因 转化表型的建立或维持，或 恶性肿瘤的进展。选择性剪接的监管负责 用于产生许多细胞和病毒的致癌和非致癌形式 癌基因。因此，更好地了解其基本机制 选择性剪接调节以及该过程的保真度可能会导致 药物设计专门影响特定物质的合成 在肿瘤发生中起关键作用的蛋白质亚型。