RNA Binding Proteins in Cancer

癌症中的 RNA 结合蛋白

• 批准号: 8444646
• 负责人: Shrikant Anant
• 金额: $ 28.38万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8444646
• 关键词: 3' Untranslated Regions; Affinity; Anchorage-Independent Growth; Apoptosis; Behavior; Binding; Binding Sites; C-terminal; Cancer Model; Cause of Death; Cell Death; Cell Survival; Cells; Colon Carcinoma; Consensus; Deletion Mutation; Disease; Elements; Epidermal Growth Factor; Epithelial Cells; Gene Expression; Gene Expression Regulation; Genetic Translation; Glycine; Goals; Growth; Interleukin-8; Intestines; Lead; Malignant Neoplasms; Manuscripts; Mediating; Messenger RNA; Methods; MicroRNAs; Mitosis; Mitotic; Modification; Mus; Mutagenesis; N-terminal; Normal Cell; Nude Mice; Pathway interactions; Phenotype; Phosphorylation; Post-Transcriptional Regulation; Process; Promoter Regions; Protein Binding; Protein Overexpression; Proteins; Proto-Oncogenes; Publishing; RNA Binding; RNA Recognition Motif; RNA Sequences; RNA Stability; RNA-Binding Proteins; Ribonucleoproteins; Ribosomes; Role; Serine; Signal Pathway; Site; Threonine; Transcript; Transcriptional Activation; Translations; Tyrosine Phosphorylation Site; Ubiquitination; United States; Vascular Endothelial Growth Factors; Work; Xenograft procedure; base; cancer cell; cyclooxygenase 2; deletion analysis; human FRAP1 protein; mRNA Decay; mRNA Stability; mTOR protein; mutant; neoplastic cell; notch protein; novel; overexpression; promoter; research study; tumor; tumor xenograft; tumorigenesis

The broad goal of the project is to understand the mechanisms by which RNA binding protein RBM3 regulates gene expression at the posttranscriptional level of mRNA stability and translation in cancer cells. We have identified that overexpression of the protein causes a normal cell to undergo a transformed phenotype resulting in the cells forming tumors in immunocompromized mice. On the other hand, knockdown of RBM3 expression results in cell death due to mitotic catastrophe. RBM3 interacts with HuR and hnRNP A1, and with AU-RNA sequences to enhance mRNA stability and translation of AU-rich transcripts such as COX-2, VEGF and IL-8. In addition, RBM3 overexpression increases the activation of the mammalian target of rapamycin protein in a Notch dependent mechanism. We have also identified microRNAs regulated upon RBM3 overexpression, including downregulating one that inhibits its own expression. Our studies also suggest that RBM3 is regulated at the posttranslational levels of phosphorylation, ubiquitination and SUMOylation. Based on these observations, we propose three specific aims. In Aim 1, we will determine the mechanism by which RNA binding protein RBM3 regulates gene expression. Here, we will identify the RNA sequences that interact with RBM3. In addition, we will determine the sequenced in COX-2 3'UTR that are required for RBM3-mediated COX-2 mRNA stability and translation. We will also identify the role of HuR and hnRNP-A1 in the process. In Aim 2, we will determine the residues in RBM3 that undergo phosphorylation, ubiquitination and SUMOylation by mutagenesis. We will also determine the effect of the mutants on mRNA stability and translation. In Aim 3, we propose to determine the mechanisms by which RBM3 induces tumorigenesis. We will use an xenograft cancer model to determine the role of mTOR and Notch pathway in RBM3 mediated tumorigenesis. Also, the role of microRNAs in the tumorigenesis will be determined. Completion of these experiments should give us a better understanding of how the RNA binding protein RBM3 functions in normal epithelial cells, and whether changes in the RBM3 expression that is observed in tumor cells is responsible for tumor behavior.

该项目的总体目标是了解 RNA 结合的机制 蛋白 RBM3 在 mRNA 转录后水平调节基因表达 癌细胞中的稳定性和翻译。我们已经发现过度表达 蛋白质导致正常细胞发生转化表型，从而产生细胞 在免疫功能低下的小鼠中形成肿瘤。另一方面，RBM3的敲低 表达导致细胞因有丝分裂灾难而死亡。 RBM3 与 HuR 相互作用并且 hnRNP A1，并具有 AU-RNA 序列，可增强 mRNA 稳定性和翻译 富含 AU 的转录物，例如 COX-2、VEGF 和 IL-8。此外，RBM3过表达 增加Notch中雷帕霉素蛋白的哺乳动物靶标的激活 依赖机制。我们还鉴定了受 RBM3 调节的 microRNA 过度表达，包括下调抑制其自身表达的表达。我们的 研究还表明 RBM3 在翻译后水平受到调节 磷酸化、泛素化和SUMO化。根据这些观察，我们 提出三个具体目标。在目标 1 中，我们将确定 RNA 的机制 结合蛋白 RBM3 调节基因表达。在这里，我们将鉴定RNA 与 RBM3 相互作用的序列。此外，我们将确定排序 COX-2 3'UTR 是 RBM3 介导的 COX-2 mRNA 稳定性所必需的 翻译。我们还将确定 HuR 和 hnRNP-A1 在此过程中的作用。在目标 2 中， 我们将确定 RBM3 中经历磷酸化、泛素化和 通过诱变进行 SUMO 化。我们还将确定突变体对 mRNA 稳定性和翻译。在目标 3 中，我们建议通过以下方式确定机制： RBM3 诱导肿瘤发生。我们将使用异种移植癌症模型 确定 mTOR 和 Notch 通路在 RBM3 介导的肿瘤发生中的作用。还， microRNA在肿瘤发生中的作用将被确定。完成这些 实验应该让我们更好地了解 RNA 结合蛋白如何 RBM3 在正常上皮细胞中发挥作用，以及 RBM3 表达是否发生变化 在肿瘤细胞中观察到的这一现象是肿瘤行为的原因。