Role of Zfp318 in the functional development of B cells

Zfp318 在 B 细胞功能发育中的作用

• 批准号: 8990957
• 负责人: Janis J. Weis
• 金额: $ 18.63万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8990957
• 关键词: Address; Affinity; Alleles; Alternative Splicing; Androgen Receptor; Animals; Antibodies; Antigens; Autoantibodies; Autoimmune Responses; B-Cell Development; B-Lymphocytes; Binding; Binding Proteins; Binding Sites; Biological Assay; Cell Lineage; Cell Maturation; Cell physiology; Chromatin; Code; Complex; DNA; DNA Binding; Data; Development; Drops; Event; Exons; Family member; Fever; Gene Expression Profile; Generations; Genes; Genetic Enhancer Element; Goals; Grant; Health; Heterogeneous Nuclear RNA; Human; Immune; Immune response; Immunization; Immunoglobulin D; Immunoglobulin M; Immunologic Deficiency Syndromes; Infection; Manuscripts; Mature B-Lymphocyte; Memory B-Lymphocyte; Messenger RNA; Modification; Molecular; Molecular Profiling; Mus; Nuclear; Pathway interactions; Precipitation; Production; Proteins; RNA; RNA Splicing; RNA-Binding Proteins; Receptor Signaling; Recurrence; Regulation; Research; Ribonucleoproteins; Role; Site; Specificity; Staging; Structure; Structure of germinal center of lymph node; Syndrome; T-Lymphocyte; Transcript; Transcriptional Activation; Transgenic Animals; Translations; United States National Institutes of Health; Vertebrates; Zinc Fingers; base; design; gene product; gene repression; immune activation; mRNA Stability; novel; promoter; protein function; research study; response; transcription factor; transcription termination

 DESCRIPTION (provided by applicant): The primary focus of this revised R21 application is on defining the functions(s) of the zinc finger family member Zfp318 in the development and function of B cells. Previously we demonstrated that Zfp318 is expressed during B cell maturation and shows the highest level of expression in mature, naïve follicular B cells. Zfp318 possesses 2 zinc finger domains as well as nuclear localization motifs: it is in a class of protein with RNA and/or DNA binding potential. We have created a Zfp318 conditional deletion allele and show in data included in this application and a manuscript in press that B cells lacking the protein fail to produce IgD. IgD, in naïve maturing and mature B cells, is coordinately expressed with IgM via alternative splicing of the Ighm/Ighd locus. B cells lacking Zfp318 continue to express VDJ-IgM but lose the ability to generate the VDJ-IgD product. New data generated since the first submission demonstrates that Zfp318 does not function to alter the alternative splicing of the Ighm/Ighd hnRNA, but instead allows for the production of this entire hnRNA by blocking the transcription termination site at the end of the IgM- encoding exons. In the absence of Zfp318, virtually all of the Ighm transcripts terminate prior to the IgD-encoding exons. Amazingly, the Ighm/Ighd gene is the only locus in B cells that shows such altered expression profiles which means that Zfp318 has evolved to singularly control this event. The experimental plan of this application section is focused upon accomplishing three specific goals. First, we will determine if there are functional consequences to the B cell and the immune response created by the absence of Zfp318. IgD and Zfp318 have co-evolved since the development of primitive vertebrates, our Zfp318 deficient animal will provide us a unique vantage to ascertain the importance of these proteins to the immune response. Secondly we will address how Zfp318 functions. Does it bind to DNA, does it bind to RNA? The fact Zfp318 only modulates Ighm/Ighd expression indicates the protein possesses a very high level of sequence/structure specificity. And finally we will address how the regulation of the expression of the Zfp318 gene is integrated into B cell development and activation. We know from our previous studies that the expression of Zfp318 is dramatically influenced by Mef2c, a transcription factor known to modulate B cell development and activation. How Zfp318 fits into this network of expression and function is the primary goal of this last question.

 描述（由应用程序提供）：此修订后的R21应用程序的主要重点是定义锌指家族成员ZFP318在B细胞开发和功能中的功能。以前，我们证明ZFP318在B细胞成熟过程中表达，并显示出成熟，幼稚的卵泡B细胞中最高水平的表达。 ZFP318具有2个锌指域以及核定位基序：它在具有RNA和/或DNA结合势的一类蛋白质中。我们已经创建了一个ZFP318条件缺失等位基因，并显示了本应用程序中包含的数据中显示的，在Press中，B细胞缺乏蛋白质无法产生IgD。在天真的成熟和成熟的B细胞中，IgD通过IGM/IGHD基因座的替代剪接与IgM协调表达。缺乏ZFP318的B细胞继续表达VDJ-IGM，但失去了生成VDJ-IGD产物的能力。自第一次提交以来生成的新数据表明，ZFP318不起作用来改变IGHM/IGHD HNRNA的替代剪接，而是允许通过在IgM-编码外显子的末端阻止转录终止位点来生产整个HNRNA。在没有ZFP318的情况下，实际上所有的IGHM转录本在IGD编码外显子之前终止。令人惊讶的是，IGHM/IGHD基因是B细胞中唯一显示出这种变化表达曲线的基因座，这意味着ZFP318已经演变为单一控制此事件。本应用部分的实验计划集中于实现三个特定目标。首先，我们会的 确定B细胞是否存在功能后果以及由于没有ZFP318而产生的免疫冲源。自从原始脊椎动物的发展以来，IGD和ZFP318已经共同发展，我们的ZFP318不足动物将为我们提供独特的Vantage，以确定这些蛋白质对免疫响应的重要性。其次，我们将解决ZFP318的功能。它与DNA结合，它与RNA结合吗？ ZFP318仅调节IGHM/IGHD表达的事实表明蛋白质具有很高的序列/结构特异性。最后，我们将解决如何将ZFP318基因表达的调节整合到B细胞的发育和激活中。从以前的研究中我们知道，ZFP318的表达受到MEF2C的极大影响，MEF2C是一种已知调节B细胞发育和激活的转录因子。 ZFP318如何适应这个表达和功能网络是最后一个问题的主要目标。