REGULATION OF HIV-1 RNA SPLICING

描述：(改编自调查人员摘要)这一目标 该项目旨在了解艾滋病毒RNA剪接的调节 病毒基因组中的正负顺式元件及其作用 与这些元素相互作用的细胞调节蛋白。 拟议的研究将集中在细胞因素对艾滋病毒的调节上。 在包括第一和第二区域及其两侧的RNA剪接 编码HIV TAT(反式激活因子)和rev基因的外显子。第一 具体目的是定义进一步的顺式作用的RNA剪接元件 HIV第一个TAT编码外显子。研究人员将在体外使用 HIV-RNA底物剪接检测及细胞内HIV-RNA的分析 瞬时转染突变的前病毒克隆以进一步鉴定 这个元素。含突变病毒的复制表型 在剪接元件方面也将进行研究。第二个目标是 鉴定和鉴定相互作用的细胞RNA结合因子 负剪接调控元素。存在的证据 与负剪接调控有关的一个或多个因素 获得了第一个TAT编码外显子中的元件。这个因素(S) 将被表征，如果它是一个新的因子，该基因将被克隆 并进行了测序。第三个目标是定义顺式作用的积极和 第二个TAT/REV编码外显子中的负剪接元件。元素 将使用体外剪接试验和瞬时 如果新的因子与这些元件结合，基因将 被克隆和测序。

DESCRIPTION: (Adapted from investigator's abstract) The goal of this project is to understand the regulation of HIV RNA splicing in terms of positive and negative cis elements within the viral genome and the role of cellular regulatory proteins which interact with these elements.The proposed studies will focus on the regulation by cellular factors of HIV RNA splicing in the regions including and flanking the first and second coding exons of the HIV tat (transactivator) and rev genes.The first specific aim is to define further cis-acting RNA splicing elements in the HIV first tat coding exon. The investigator will use in vitro splicing assays with HIV RNA substrates and analysis of HIV RNA in cells transiently transfected with mutant proviral clones to further define this element. The replication phenotype of virus containing mutations in he splicing element will also be studied. The second aim is to identify and characterize cellular RNA-binding factors interacting with the negative splicing regulatory element. Evidence for the presence of a factor or factors that bind to the negative splicing regulatory element in the first tat coding exon has been obtained. This factor(s) will be characterized and, if it a novel factor, the gene will be cloned and sequenced. The third goal is to define cis-acting positive and negative splicing elements in the second tat/rev coding exon. Elements will be defined using in vitro splicing assays and transient transfections.If novel factors bind to these elements, the genes will be cloned and sequenced.