RNA Signaling Control in Rous Sarcoma Virus

劳斯肉瘤病毒中的 RNA 信号传导控制

• 批准号: 6681255
• 负责人: MARK T MCNALLY
• 金额: $ 28.2万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-14 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6681255
• 关键词: RNA splicing; Rous sarcoma virus; crosslink; gel mobility shift assay; genetic regulatory element; intermolecular interaction; polyadenylate; posttranscriptional RNA processing; small nuclear ribonucleoproteins; tissue /cell culture; transcription factor; ultraviolet radiation; virus genetics; virus replication

DESCRIPTION (provided by applicant): As a model to study regulated RNA processing of pre-mRNAs in eukaryotic cells, we are studying viral cis elements and host trans-acting factors required for proper RNA processing and replication of Rous sarcoma virus (RSV). RSV primary transcripts are spliced and polyadenylated by the host RNA processing machineries to generate env/src mRNA, but the process must be controlled since the virus requires that a majority (75%) of the RNA remain unspliced to serve as mRNA for the gag-pol proteins and as genomic RNA for progeny virions. A novel RNA element (the NRS) that binds numerous splicing factors is required for accumulation of appropriate unspliced RNA levels and also for proper polyadenylation. NRS mutations can lead to replication defects due to oversplicing and to pathogenesis from the increased production of poly(A) read-through transcripts that play a role in oncogenesis via insertional activation. Aim I will investigate the role of various factors in NRS splicing inhibition and explore mechanisms of inhibition involving novel, inappropriate snRNP interactions. In Aim II, cis elements and trans factors that mediate efficient snRNP binding to the NRS will be studied. In Aim lll, the role of the NRS in promoting efficient polyadenylation will be investigated. In Aim IV, the function of a second splicing suppressor (SSS) associated with the src 3' splice site will be studied. These Aims are significant in that they will enhance our understanding of control of critical RNA processing steps required for viral replication and pathogenesis, and the RSV system provides a powerful tool for dissecting novel cellular mechanisms of RNA processing regulation.

描述（申请人提供）：作为研究真核细胞中前体 mRNA 受调节 RNA 加工的模型，我们正在研究劳斯肉瘤病毒 (RSV) 正确 RNA 加工和复制所需的病毒顺式元件和宿主反式作用因子。 RSV 初级转录本由宿主 RNA 加工机器进行剪接和聚腺苷酸化，以生成 env/src mRNA，但该过程必须受到控制，因为病毒需要大多数（75%）的 RNA 保持未剪接状态，以充当 gag 的 mRNA。 pol 蛋白和作为子代病毒颗粒的基因组 RNA。需要一种结合多种剪接因子的新型 RNA 元件（NRS）来积累适当的未剪接 RNA 水平以及适当的聚腺苷酸化。 NRS 突变可能因过度剪接而导致复制缺陷，也可能因 Poly(A) 通读转录物产量增加而导致复制缺陷，而 Poly(A) 通读转录物通过插入激活在肿瘤发生中发挥作用。目标我将研究各种因素在 NRS 剪接抑制中的作用，并探索涉及新的、不适当的 snRNP 相互作用的抑制机制。在目标 II 中，将研究介导 snRNP 与 NRS 有效结合的顺式元件和反式元件。在目标III中，将研究NRS在促进有效聚腺苷酸化中的作用。在目标 IV 中，将研究与 src 3' 剪接位点相关的第二剪接抑制子 (SSS) 的功能。这些目标意义重大，因为它们将增强我们对病毒复制和发病机制所需的关键 RNA 加工步骤的控制的理解，并且 RSV 系统为剖析 RNA 加工调节的新型细胞机制提供了强大的工具。