Characterization of TRBP-containing complexes

含TRBP复合物的表征

• 批准号: 7661577
• 负责人: RAMIN SHIEKHATTAR
• 金额: $ 28.83万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7661577
• 关键词: Biochemical; Caenorhabditis elegans; Cell Nucleus; Cells; Complex; Cytoplasm; Data; Development; Developmental Process; Dicer Enzyme; Distant; Double-Stranded RNA; Enzymes; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Genome; Hematopoietic; Human; In Vitro; Individual; Introns; Link; Maintenance; Malignant Neoplasms; Mammals; Mediating; Messenger RNA; MicroRNAs; Minority; Modeling; Nucleotides; Oncogenic; Pathway interactions; Pattern; Process; Proteins; RNA Interference; RNA-Induced Silencing Complex; Recombinants; Regulation; Ribonuclease III; Ribosomes; Role; Small Interfering RNA; Small RNA; Structure; Testing; Time; Translational Repression; Translations; Untranslated Regions; Work; ds RNA-Binding Proteins; gel electrophoresis; human DICER1 protein; in vivo; insight; neuron development; neuronal patterning; polypeptide; pre-miRNA; pri-miRNA; protein protein interaction; research study; response; stem; Biochemical; Caenorhabditis elegans; Cell Nucleus; Cells; Complex; Cytoplasm; Data; Development; Developmental Process; Dicer Enzyme; Distant; Double-Stranded RNA; Enzymes; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Genome; Hematopoietic; Human; In Vitro; Individual; Introns; Link; Maintenance; Malignant Neoplasms; Mammals; Mediating; Messenger RNA; MicroRNAs; Minority; Modeling; Nucleotides; Oncogenic; Pathway interactions; Pattern; Process; Proteins; RNA Interference; RNA-Induced Silencing Complex; Recombinants; Regulation; Ribonuclease III; Ribosomes; Role; Small Interfering RNA; Small RNA; Structure; Testing; Time; Translational Repression; Translations; Untranslated Regions; Work; ds RNA-Binding Proteins; gel electrophoresis; human DICER1 protein; in vivo; insight; neuron development; neuronal patterning; polypeptide; pre-miRNA; pri-miRNA; protein protein interaction; research study; response; stem

DESCRIPTION (provided by applicant): The microRNA-group of genes of C. elegans is required to control the timing of larval development and neuronal patterning. A predominant number of these genes are conserved in mammals and recent experiments demonstrated a role for them in modulation of hematopoietic differentiation and cancer. Our biochemical characterization of microRNA pathway revealed the association of the RNase III enzyme Dicer with the oncogenic protein TRBP. Dicer is the enzyme responsible for the generation of mature microRNA and processing of exogenous double-stranded RNA to short interfering RNAs (siRNAs). Biochemical analysis of TRBP-containing complexes not only confirmed the association of TRBP and Dicer but also uncovered the presence of Argonaute 2, the catalytic engine of RNA-induced silencing complex as a stable component of Dicer/TRBP complex. We hypothesize that these three proteins form the core of a complex responsible for processing and execution of RNAi response. We also uncovered evidence indicating that the 60S ribosome subunit associates with TRBP to form a complex involved in mediating microRNA-directed translational repression. Aim 1 will define the function of Dicer/TRBP complex in the genesis of siRNA/miRNA and begin to examine the role of individual proteins and their domain structure in the regulation of siRNA/miRNA processing activity. Aim 2 describes the detailed functional analysis of the association of Dicer/TRBP complex with the effector complex containing the Ago2 subunit and delineates the role of individual proteins in the functioning of the posttranscriptional gene silencing. Aim 3 extends the work to the analysis of the large TRBP- containing complex, which is associated with the 60S ribosome subunit and the human Armitage protein involved in translational repression.

描述（由申请人提供）：秀丽隐杆线虫基因的microRNA基因需要控制幼虫发育和神经元模式的时间。这些基因的主要数量在哺乳动物中是保守的，最近的实验证明了它们在调节造血分化和癌症中的作用。我们对microRNA途径的生化表征揭示了RNase III酶DICER与致癌蛋白TRBP的关联。 DICER是负责产生成熟microRNA的酶，并处理外源性双链RNA到短干扰RNA（siRNA）。对含TRBP的复合物的生化分析不仅证实了TRBP和DICER的关联，而且还发现了Argonaute 2的存在，Argonaute 2是RNA诱导的沉默复合物的催化引擎，是DICER/TRBP复合物的稳定成分。我们假设这三种蛋白质构成了负责处理和执行RNAi反应的复合物的核心。我们还发现了证据，表明60年代的核糖体亚基与TRBP相关，以形成参与介导microRNA定向翻译抑制的复合物。 AIM 1将定义DICER/TRBP复合物在siRNA/miRNA的起源中的功能，并开始检查单个蛋白质及其结构结构在siRNA/miRNA加工活性调节中的作用。 AIM 2描述了DICER/TRBP复合物与含有AGO2亚基的效应子复合物的关联的详细功能分析，并描绘了单个蛋白质在转录后基因沉默的功能中的作用。 AIM 3将工作扩展到对含有大型TRBP的复合物的分析，该复合物与60S核糖体亚基和参与转化抑制的人类武器蛋白有关。