REGULATION OF PRE-MRNA SPLICING

描述(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，与 致癌。基因选择性剪接调控的基本机制 细胞和病毒基因在不同组织、发育阶段或在 对外部信号的响应，将进行调查。我们将开展详细的工作 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‘它在替代物5中的活性’ 剪接位点选择、剪接沉默和端粒长度调节； 基因毒性对其定位和活性的调节 压力。我们将在培养的哺乳动物细胞中进行遗传筛选 确定与前mRNA剪接的保真度有关的蛋白质组分。 最后，我们将继续开发涉及Rational的新技术 设计模拟SR蛋白质的一种活性的化合物， 通过外显子剪接增强子激活剪接。这些化合物将是 测试他们纠正由以下原因引起的特定剪接缺陷的能力 癌症易感基因的突变，并调节其表达 另一种细胞凋亡基因的剪接异构体。这些研究是 与计划项目的总体目标直接相关。 作为替代前mRNA剪接的全球调节者，hnRNP A1及其 拮抗剂SR蛋白可能是所观察到的异常的原因 转化细胞中多个基因的mRNA表达模式。其中 这些调节器的潜在靶点是几个关键基因，涉及 建立或维持转化的表型，或在 恶性进展。对选择性剪接的监管是负责的 用于产生许多细胞和病毒的致癌和非致癌形式 致癌基因。因此，更好地理解 选择性的剪接调控，以及这一过程的保真度，可能会导致 以药物的设计来具体影响特定的合成 在肿瘤发生中起关键作用的蛋白质亚型。

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.