Biochemistry of Eukaryotic Messenger RNA Synthesis

真核生物信使 RNA 合成的生物化学

• 批准号: 8294646
• 负责人: JOAN W CONAWAY
• 金额: $ 38.96万
• 依托单位: STOWERS INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8294646
• 关键词: Acute Myelocytic Leukemia; Biochemical; Biochemical Process; Biochemistry; Chromatin; Code; Collection; Complex; DNA Polymerase II; Elongation Factor; Enzymes; Etiology; Family member; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Human; In Vitro; Investigation; Light; Malignant Neoplasms; Malignant neoplasm of liver; Mammalian Cell; Messenger RNA; Molecular; Oncogenes; Polymerase; Primary carcinoma of the liver cells; Proteins; RNA Polymerase II; RNA chemical synthesis; Regulation; Research; Role; Site; System; Transcript; Transcription Elongation; Transcription Initiation; UV induced; Ubiquitination; base; elongin; human disease; insight; novel; promoter; public health relevance; reconstitution; transcription factor

DESCRIPTION (provided by applicant): Our long range goal is to contribute to an understanding of the biochemical mechanisms underlying the regulation of eukaryotic messenger RNA synthesis. The major goal of the research described in this proposal is to determine, at the molecular level, how promoter-specific transcription by RNA polymerase II (Pol II) is controlled by the action of a set of transcription factors that direct accurate initiation and efficient elongation of transcripts in vitro. Our overall approach is essentially the same as that described in the previous proposals: (i) to reconstitute in vitro, with purified proteins, faithful and efficient transcription of eukaryotic protein-coding genes and (ii) to use this reconstituted system to elucidate the biochemical mechanisms governing transcription initiation and elongation by mammalian Pol II. This research will be organized along the following lines: (1) Further analysis of the mechanism of action of Pol II elongation factor Elongin A and of its newly described role in UV-induced ubiquitination of Pol II. (2) Further investigation of the mechanism of action of Pol II elongation factor ELL. (3) Detailed analysis of the mechanisms of action of two novel human ATP-dependent chromatin remodelers referred to as the human INO80 complex and the SNF2 family member and oncogene ALC1 (Activated in Liver Cancer 1). These studies should provide information on the basic mechanisms governing eukaryotic messenger RNA synthesis, and they also have the potential to yield new insights into the molecular bases of cancers resulting from abherrant expression of ELL and its associated proteins and of ALC1. PUBLIC HEALTH RELEVANCE: It is well established that eukaryotic messenger RNA synthesis is a major site for the regulation of gene expression and, further, that its misregulation often results in human diseases, including many forms of cancer. Eukaryotic messenger RNA synthesis is a complex biochemical process catalyzed by the enzyme RNA polymerase II with the assistance of a diverse collection of transcription factors. Despite considerable progress over the past 20 years, critical aspects of the mechanisms underlying misregulation of eukaryotic messenger RNA synthesis in human disease are not completely understood. As a consequence, the goal of the research described in this proposal is to contribute to an understanding of the biochemical mechanisms underlying the regulation of messenger RNA synthesis in mammalian cells. In particular, we plan to pursue investigations of the mechanisms of action of several transcription factors discovered in our lab. In addition to shedding light on the basic mechanisms governing mammalian messenger RNA synthesis, these studies have the potential to shed light on aspects of cancer, since two transcription factors, ELL and ALC1, which we plan to investigate have been proposed to participate in the etiology of acute myeloid leukemia and hepatocellular carcinoma, respectively.

描述（由申请人提供）： 我们的长期目标是促进对真核生物信使RNA合成调控的生化机制的理解。本提案中描述的研究的主要目标是在分子水平上确定RNA聚合酶II（Pol II）的启动子特异性转录如何受一组转录因子的作用控制，这些转录因子在体外指导转录物的准确起始和有效延伸。我们的整体方法基本上是相同的，在以前的建议中所描述的：（i）在体外重建，与纯化的蛋白质，忠实和有效的真核生物蛋白质编码基因的转录和（ii）使用这个重建系统来阐明的生物化学机制，转录起始和延长哺乳动物Pol II。本研究将沿着以下路线组织：（1）进一步分析Pol II延伸因子Elongin A的作用机制及其在UV诱导的Pol II泛素化中的新作用。(2)进一步研究Pol II延伸因子ELL的作用机制。(3)详细分析了两种新型人类ATP依赖性染色质重塑剂（称为人类INO80复合物和SNF2家族成员）和癌基因ALC 1（在肝癌中活化1）的作用机制。 这些研究应该提供真核生物信使RNA合成的基本机制的信息，他们也有可能产生新的见解，从异常表达的ELL及其相关蛋白和ALC 1的癌症的分子基础。 公共卫生相关性： 已经确定，真核信使RNA合成是基因表达调控的主要位点，并且进一步地，其失调经常导致人类疾病，包括许多形式的癌症。真核生物信使RNA合成是一个复杂的生物化学过程，由RNA聚合酶II在多种转录因子的协助下催化。尽管在过去的20年中取得了相当大的进展，但人类疾病中真核信使RNA合成失调的机制的关键方面尚未完全了解。因此，本提案中描述的研究目标是有助于理解哺乳动物细胞中信使RNA合成调控的生化机制。特别是，我们计划继续研究我们实验室发现的几种转录因子的作用机制。除了阐明哺乳动物信使RNA合成的基本机制外，这些研究还有可能阐明癌症的各个方面，因为我们计划研究的两种转录因子ELL和ALC1分别参与了急性髓细胞白血病和肝细胞癌的病因。