Novel Subunit of NF-kB Confers Gene Regulatory Specificity

NF-kB 的新亚基赋予基因调控特异性

基本信息

批准号：
8210985
负责人：
Fengyi Wan
金额：
$ 24.15万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-05 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8210985
关键词：
20 year old Address Affinity Affinity Chromatography Apoptosis Award B-Lymphocytes Binding Binding Sites Biochemical Biochemical Pathway Biological Assay Cell Line Cell Nucleus Cell physiology Cells Chronic Complex Cytoplasm DNA DNA Binding DNA Sequence Data Disease Electrophoretic Mobility Shift Assay Environment Evolution Faculty Gene Expression Gene Expression Profile Gene Expression Regulation Gene Targeting Genes Genetic Transcription Goals Heterogeneous-Nuclear Ribonucleoprotein K Human IL2RA gene Immune response Immunoglobulins In Vitro Inflammation Inflammatory Ink Instruction Interleukin-2 Intervention KH Domain Knowledge Length Ligands Light Link Malignant Neoplasms Mammalian Cell Maps Mass Spectrum Analysis Mediating Mentorship Microarray Analysis Modeling Molecular Mus NF-kappa B Nuclear Extract Pathway interactions Peptides Peripheral Blood Lymphocyte Phase Physiological Physiological Processes Play Positioning Attribute Proteins Public Health Recombinants Regulation Regulator Genes Reporter Genes Research Research Personnel Role Sequence Analysis Signal Pathway Signal Transduction Site Specificity Superantigens T-Cell Activation T-Cell Lymphoma T-Lymphocyte TNFRSF5 gene Tertiary Protein Structure Therapeutic Therapeutic Intervention Time Transcriptional Regulation Universities Viral Genes Work base cancer therapy career development cell growth cell type chromatin immunoprecipitation design dimer drug discovery empowered experience improved in vivo in vivo Model inhibitor/antagonist innovation insight intercellular communication model design new therapeutic target novel p65 reconstitution research study response ribosomal protein S3 skills small molecule synaptotagmin I transcription factor

项目摘要

The transcription factor NF-kB extensively regulates a plethora of genes and is involved in cancer and inflammatory disease. Aberrant constitutive NF-kB activation has been recognized as a critical pathogenetic in cancer and inflammatory disease. Our long-term goal is to elucidate the mechanism controlling specific expression of NF-kB target genes. The specific hypothesis is that NF-kB DNA binding complexes contain novel components, besides the Rel dimer and these mediate specific gene regulation. We based that hypothesis on the observations 1) the original size estimate of native NF-kB in nuclear extracts was > 200 kD, but when reconstituted from purified p50 and p65 proteins, it was estimated to be 115 kD which would be appropriate for a simple heterodimer, 2) reconstituted heterodimers from purified proteins have a > 100 fold lower affinity than native NF-kB, 3) our recent finding that ribosomal protein S3 (RPS3) is a novel component in NF-kB DNA binding complexes that mediates selective gene regulation. The experimental focus of this proposal is to identify the novel subupjtsgfNFTkB andsto define the biochemical function of rion-Rgl ^ suburiits. The specific aims are: 1. to elucidate the mechanism of RPS3-mediated specificity in NF-kB target gene regulation; 2. to identify novel component(s) in NF-kB DNA binding complexes that mediated specific gene expression. We will purify the novel components in CD25 kB DNA binding complex, elucidate the functional significance and mechanism of novel components in NF-kB signaling under normal and pathological settings; 3, to develop in vivo models and design novel small peptides targeting RPS3 and novel components in NF-kB DNA binding complexes to specifically irihibitor NF-kB genes. We will map the minimal parts in p65, RPS3 and novel components for physical interaction and NF-kB function and assess the inhibitory capability of cell-permeable peptides targeting at these minimal parts. This project will provide new insight into NF-kB and better our understanding of the regulatory specificity of NF-kB. These results will also provide potential novel therapeutic targets for the treatment of cancer and inflammatory disease, and may have implication for NF-kB-related diseases in general.

转录因子NF-KB广泛调节了许多基因，并参与癌症和炎症性疾病。异常的本构NF-KB激活已被认为是一种关键的致病性在癌症和炎症性疾病中。我们的长期目标是阐明控制特定的机制 NF-KB靶基因的表达。具体假设是NF-KB DNA结合复合物包含新成分，除了相关二聚体和这些介导的特定基因调节。我们以此为基础关于观察结果的假设1）核提取物中天然NF-KB的原始大小估计值> 200 KD，但是当从纯化的P50和P65蛋白中重构时，估计为115 kd 适用于简单的异二聚体，2）纯化蛋白质的重构异二聚体具有> 100倍比天然NF-KB低的亲和力，3）我们最近发现核糖体蛋白S3（RPS3）是一种新成分的发现在NF-KB DNA结合复合物中介导选择性基因调节的复合物。实验重点建议是识别新型subupjtsgfnftkb，并定义rion-rgl ^的生化功能郊区。具体目的是：1。阐明NF-KB目标中RPS3介导的特异性的机制基因调节； 2。确定NF-KB DNA结合复合物中的新成分，介导了特定基因表达。我们将纯化CD25 Kb DNA结合复合物中的新成分，阐明 NF-KB信号在正常和病理环境； 3，开发体内模型并设计针对RPS3和新颖的新型小肽 NF-KB DNA结合复合物中的成分与特异性irihibitor NF-KB基因。我们将绘制最小的 P65，RPS3和新型组件的零件，用于物理相互作用和NF-KB功能，并评估靶向这些最小部分的可渗透肽的抑制能力。该项目将提供新的深入了解NF-KB，并更好地理解NF-KB的监管特异性。这些结果也将为治疗癌症和炎症性疾病提供潜在的新型治疗靶一般来说，对NF-KB相关疾病有影响。