ATF3-Mediated Transcriptional Repression of Interferon-beta

ATF3 介导的干扰素-β 转录抑制

• 批准号: 7883393
• 负责人: STEVEN LEE WEINSTEIN
• 金额: $ 11.48万
• 依托单位: SAN FRANCISCO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-03 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7883393
• 关键词: Acetylation; Acute; Animal Experiments; Autoimmune Diseases; Autoimmunity; Bacterial Infections; Binding; Binding Sites; Biological Assay; CCL4 gene; Cell Wall; Cell physiology; Cells; Complex; Consensus; Consensus Sequence; Cytokine Gene; Data; Development; Educational process of instructing; Exposure to; Funding; Genes; Genetic Transcription; Goals; Grant; Histone Deacetylation; Histones; Hypersensitivity; Immune response; Immune system; Infection; Inflammation; Inflammatory Response; Interferon-beta; Interferons; Investigation; JUN gene; Journals; Lead; Link; Lipopolysaccharides; Literature; Measures; Mediating; Mediator of activation protein; Messenger RNA; Molecular; Outcome; Pathology; Peer Review; Production; Productivity; Protein Binding; Proteins; Publishing; Regulation; Reporter; Reporting; Repression; Research; Research Peer Review; Rest; Scientist; Septic Shock; Simulate; Site; Source; Stimulus; Students; Testing; Time; Transactivation; Viral; Work; activating transcription factor 3; base; career; chromatin immunoprecipitation; cytokine; design; dimer; gene repression; improved; in vivo; mRNA Expression; macrophage; meetings; microbial; prevent; programs; promoter; research study; self esteem; transcription factor

DESCRIPTION (provided by applicant): The immune system mediator, IFN-?, is a critical part of the anti-viral defense and it is casually linked to lethal septic shock and some autoimmune diseases. Increased understanding of how IFN-? is regulated by our cells is a key step toward creating better remedies to treat septic shock and autoimmunity. This investigation, designed to yield new information about the regulation of IFN-?, contributes to that goal. Acute inflammation is an early immune response to infection that aids in microbial clearance. Macrophages (Mo) at the infected site trigger the inflammatory response by secreting cytokines. The long-term objective of this research is to understand the molecular mechanisms that regulate the production of cytokines by Mo. Interferon-beta (IFN-?) is a cytokine produced by Mo and other cells that functions in anti-viral defense as well as modulates immune responses that cause autoimmunity. Expression of IFN-? in Mo is primarily controlled transcriptionally. In resting Mo, transcription of IFN-? is repressed, but upon infection, mRNA synthesis is induced. Many of the proteins that bind to the IFN-? promoter and transactivate this gene are well known, while the components that repress transcription are less well characterized. Recently, we discovered that the transcriptional regulator, Activating Transcription Factor 3 (ATF-3), represses IFN-? transcription. Mo lacking ATF-3 or expressing reduced amounts of ATF-3 produce higher levels of IFN-? mRNA after IPS stimulation, compared to wild type Mo. These observations lead us to hypothesize that ATF-3 binds to the IFN-? promoter and represses transcription of this gene in Mo. The specific aims are designed to test this hypothesis by: (1) determining whether over-expression of ATF-3 in Mo represses IFN-? transcription; and (2) determining whether ATF-3 binds to the IFN-¿ promoter; (3) determining whether ATF-3-mediated repression of IFN-? transcription is caused by histone deacetylation. To test for ATF-3-mediated repression of IFN-? transcription, ATF-3 will be over-expressed and IFN-? transcription will be measured in a reporter assay. In vivo binding of ATF-3 to the IFN-? promoter will be tested by chromatin immunoprecipitation (ChlP). The possibility that repression of IFN-? transcription by ATF-3 is caused by histone deacetylation will be tested by examining the acetylation state of IFN-? promoter histones in the presence or absence of ATF-3 by ChlP.

性状（由申请方提供）：免疫系统介质IFN-？，是抗病毒防御的关键部分，它与致命的败血性休克和一些自身免疫性疾病有着偶然的联系。增加了解如何干扰素-？是创造更好的治疗感染性休克和自身免疫的方法的关键一步。本研究旨在获得有关IFN-？调节的新信息，有助于实现这一目标。急性炎症是对感染的早期免疫反应，有助于微生物清除。感染部位的巨噬细胞（Mo）通过分泌细胞因子触发炎症反应。本研究的长期目标是了解Mo调节细胞因子产生的分子机制。干扰素-β（IFN-？）是由Mo和其他细胞产生的细胞因子，其在抗病毒防御中起作用，并调节引起自身免疫的免疫应答。IFN-γ的表达在Mo中主要是转录控制的。在静息状态下，IFN-？被抑制，但在感染时，mRNA合成被诱导。许多与IFN-？启动子和反式激活该基因是众所周知的，而抑制转录的组分还不太清楚。最近，我们发现，转录调节因子，激活转录因子3（ATF-3），抑制IFN-？转录。Mo缺乏ATF-3或表达ATF-3量减少产生更高水平的IFN-？IPS刺激后的mRNA，与野生型Mo.这些观察使我们假设，ATF-3结合IFN-？启动子并抑制该基因在Mo.具体的目的是为了检验这一假设：（1）确定是否过表达ATF-3在Mo抑制IFN-？转录;（2）确定ATF-3是否与IFN-γ启动子结合;（3）确定ATF-3是否介导IFN-γ的抑制？转录是由组蛋白脱乙酰化引起的。测试ATF-3介导的抑制IFN-？转录时，ATF-3会过表达，而IFN-？转录将在报告基因测定中测量。ATF-3与IFN-？将通过染色质免疫沉淀（ChIP）测试启动子。抑制IFN-？ATF-3转录是由组蛋白去乙酰化引起的，将通过检查IFN-γ的乙酰化状态来检测。ChlP在存在或不存在ATF-3的情况下启动子组蛋白。