Determinants of BATF Function

BATF功能的决定因素

• 批准号: 8635120
• 负责人: ELIZABETH J TAPAROWSKY
• 金额: $ 18.37万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-02 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8635120
• 关键词: Address; Allergic Disease; Antibody Formation; Apoptotic; Autoimmunity; B-Lymphocytes; Behavior; Binding; Biological; Biological Assay; CD4 Positive T Lymphocytes; Cell physiology; Cells; Communicable Diseases; Complex; Consensus; Critical Pathways; DNA; DNA Binding; Data; Development; Dimerization; Disease; Drug Design; Elements; Event; Family; Family member; Foundations; Future; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Health; Helper-Inducer T-Lymphocyte; Homeostasis; Homing; Human; IL17 gene; IRF4 gene; Immune; Immune System Diseases; Immune response; Immune system; Inflammation; Knowledge; Laboratories; Learning; Light; Lymphocyte; Lymphoproliferative Disorders; Mediating; Mission; Modeling; Modification; Molecular; Mus; Mutation; Nuclear; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Positioning Attribute; Post-Translational Protein Processing; Prevention; Process; Property; Protein Engineering; Protein Family; Proteins; Proto-Oncogene Proteins c-jun; Public Health; Receptor Gene; Recruitment Activity; Regulation; Regulator Genes; Reporter Genes; Retinoic Acid Receptor; Role; Site; Staging; T-Lymphocyte; Testing; Transcription Factor AP-1; Transgenic Organisms; Up-Regulation; Variant; Work; design; dimer; extracellular; gene repression; human CREB3 protein; improved; in vivo; interest; jun Oncogene; member; novel therapeutics; pathogen; preference; prevent; protein function; public health relevance; retroviral-mediated; small molecule; tool; transcription factor

PROJECT SUMMARY BATF is the founding member of the BATF family of basic leucine zipper proteins and a major contributor to the function of AP-1 transcription factor complexes in immune cells. Batf null (Batf KO) mice are viable, yet display a number of phenotypes that position the protein within pathways critical for the regulation of inflammation, autoimmunity, host protection against pathogens and the prevention of lymphoproliferative disorders. As a result, there is enormous interest in using BATF, its regulators, and its effector genes in targeted strategies to control these health concerns. However, in order to assess which strategy to pursue, it is critical to identify targetable determinants that control BATF function with an emphasis on those determinants that discriminate between BATF function in different cellular contexts. This proposal is designed to address this gap in our knowledge. What is known is that BATF forms heterodimers with the JUN AP-1 proteins and that JUN:BATF dimers regulate genes by binding classic AP-1 DNA, or by recruiting IRF transcription factors to composite DNA elements called AICE. Both target gene activation and repression are associated with BATF expression in cells, but the precise manner by which these functions are distinguished has not been fully described. Additionally, there is evidence that BATF proteins can impact gene expression in the absence of DNA binding and that two phosphorylation events alter BATF target gene regulation by impacting its DNA binding and dimerization properties. The goal of this proposal is to test the hypothesis that phosphorylation of BATF selectively modulates BATF protein function in vivo. In Aim 1, BATF proteins engineered to mimic phosphorylation events and containing additional mutations that disrupt association with IRF4 and EGR2, will be assayed for DNA binding properties and for the ability to influence transcription using both AP-1 and AICE regulated genes. In Aim 2, the behaviors characterized in Aim 1 will be tested for their function in three, different, Batf-dependent transcription programs by examining BATF variants for their ability to rescue Batf KO T cell phenotypes. Completion of these aims will address our immediate goals of defining how naturally-occurring post-translational modifications impact the molecular properties of BATF and influence their competency to direct specific T cell functions. Results from this R21 will set the stage for future work to identify the kinase/phosphatase network(s) directing these BATF modifications in vivo and to profile how the cellular transcriptome responds following BATF modification. Reversible phosphorylation is among the most successful approaches for targeted drug design. A small molecule approach aimed at manipulating BATF post-translational modifications to inhibit, or augment, BATF function in vivo, will be a valuable tool for the management of immune system disorders.

项目摘要 BATF是碱性亮氨酸拉链蛋白BATF家族的创始成员， 免疫细胞中AP-1转录因子复合物的功能。Batf null（Batf KO）小鼠是可行的，但 显示出许多表型，这些表型将蛋白质定位在对调节 炎症、自身免疫、宿主对病原体的保护和淋巴增生性疾病的预防 紊乱因此，人们对使用BATF、其调节因子和其效应基因在 有针对性的战略来控制这些健康问题。然而，为了评估采取何种战略， 关键是确定控制BATF功能的目标决定因素，重点是 决定因素区分不同细胞环境中的BATF功能。该提案旨在 来填补我们知识上的空白。已知的是BATF与JUN AP-1形成异二聚体 JUN：BATF二聚体通过结合经典的AP-1 DNA或通过募集IRF来调节基因， 转录因子合成的DNA元件称为AICE。靶基因的激活和抑制都是 与BATF在细胞中的表达相关，但这些功能区分的精确方式 还没有完全描述。此外，有证据表明，BATF蛋白可以影响基因表达， DNA结合缺乏和两个磷酸化事件改变了BATF靶基因调节， 影响其DNA结合和二聚化性质。本提案的目的是检验以下假设： BATF的磷酸化在体内选择性地调节BATF蛋白功能。在目标1中，BATF蛋白 工程化以模拟磷酸化事件，并含有破坏与 IRF 4和EGR 2的DNA结合特性和影响转录的能力将使用 AP-1和AICE调节基因。在目标2中，将测试目标1中描述的行为， 通过检查BATF变体的能力， 拯救Batf KO T细胞表型。完成这些目标将有助于实现我们的近期目标， 自然发生的翻译后修饰如何影响BATF的分子特性， 它们指导特定T细胞功能的能力。R21的结果将为未来的工作奠定基础， 鉴定在体内指导这些BATF修饰的激酶/磷酸酶网络，并分析 细胞转录组在BATF修饰后响应。可逆的磷酸化是最重要的 靶向药物设计的成功方法。一种旨在操纵BATF的小分子方法 翻译后修饰以抑制或增强体内BATF功能，将是研究BATF的有价值的工具。 免疫系统疾病的管理。