Studies of Antibody Diversity and Genome Stability: Regulation of Activation-Indu

抗体多样性和基因组稳定性的研究：激活诱导的调节

• 批准号: 8409854
• 负责人: Yee Ling Wu
• 金额: $ 0.79万
• 依托单位: MEDICAL RES COUNCIL LAB OF MOLEC BIOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-11-24 至 2013-11-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8409854
• 关键词: Address; Antibodies; Antibody Affinity; Antibody Diversity; Antigens; B-Lymphocytes; Biochemical; Cancer Etiology; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromosomal translocation; Co-Immunoprecipitations; Complex; Coupled; DNA; DNA Sequence; DNA repair protein; Data; Deamination; Deoxycytidine; Disease; Embryo; Ensure; Enzyme Activation; Enzymes; Equilibrium; Event; Fellowship; Gene Conversion; Gene Expression; Generation of Antibody Diversity; Generations; Genes; Genetic Transcription; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Hereditary Disease; Immune; Immunoglobulin Class Switching; Immunoglobulin Constant Region; Immunoglobulin Somatic Hypermutation; Immunoglobulin Switch Recombination; Immunoglobulins; Immunologics; Immunoprecipitation; Induced Mutation; Infection; Knockout Mice; Knowledge; Lesion; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microbe; Molecular; Molecular Profiling; Mus; Mutate; Mutation; Normal Cell; Organism; Outcome; Physiological; Process; Proteins; Proteomics; Proto-Oncogenes; RNA Splicing; Regulation; Specificity; Tertiary Protein Structure; Tumor Suppressor Genes; Uracil; Vertebrates; abstracting; activation-induced cytidine deaminase; cis acting element; immune function; insight; leukemia/lymphoma; microbial; mutant; next generation; novel; novel therapeutics; pathogen; protein activation; research study

Abstract: Activation-induced deaminase (AID) is an enzyme that mutates genes for antibodies in vertebrates. These AID-mediated mutations are necessary for an organism to generate specific antibodies to recognize and eradicate great varieties of microbial infections. However, if AID is not properly regulated, it can change DNA sequences of genes important for normal cell functions and cause diseases. Indeed, some lymphomas and leukemias are caused by abnormal activities of AID. How AID can change DNA sequences only in antibody genes but not in other genes is poorly understood. Recently, the catenin ¿ like 1 protein (CTNNBL1) has been found to physically interact with AID. Little is known for the physiologic function of CTNNBL1. However, the interaction with CTNNBL1 is required for AID to change DNA sequences specifically in the antibody genes. In this study, a combination of high throughput approaches including next-generation sequencing and microarray analyses and targeted biochemical and immunologic analyses will be used to investigate how AID specifically mutate antibody genes, particularly what genomic regions are accessible to AID, both in the presence and in the absence of CTNNBL1. These experiments will also detect any alterations in transcription and splicing caused by the absence of CTNNBL1, and thereby shed lights on normal cellular functions of CTNNBL1. Mass spectrometry, mutagenic analyses and immunologic experiments will be performed to determine the molecular details of the interaction between AID and CTNNBL1 as well as to elucidate the mechanisms by which CTNNBL1 regulates the targeted activity of AID. It is anticipated that results from this study will be highly relevant for understanding the mechanisms underlying the generation of antibody diversity, and in the creation of new therapeutic strategies to minimize genomic disorders and cancers.

抽象的： 激活诱导的脱氨酶（AID）是一种酶，可突变脊椎动物中抗体的基因。这些 辅助介导的突变对于生物体生成特定抗体以识别和 消除微生物感染的大变化。但是，如果辅助不适当调节，则可以改变DNA 基因序列对于正常细胞功能很重要并引起疾病。确实，一些淋巴瘤和 白血病是由异常活动引起的。辅助如何仅在抗体中更改DNA序列 基因但在其他基因中没有理解。最近，catenin - 像1蛋白（CTNNBL1）一样 发现与援助物理互动。 CTNNBL1的生理功能闻名很少。但是， 与CTNNBL1的相互作用是需要在抗体基因中特异性更改DNA序列的辅助所必需的。 在这项研究中，高吞吐量方法的结合包括下一代测序和 微阵列分析和针对性的生化和免疫学分析将用于研究如何帮助 特异性突变抗体基因，尤其是在哪些基因组区域可用于帮助的地方 存在和不存在CTNNBL1。这些实验还将检测到转录的任何改变 和由CTNNBL1缺乏引起的剪接，从而在正常的细胞功能上脱光灯 ctnnbl1。将进行质谱，诱变分析和免疫学实验 确定AID和CTNNBL1之间相互作用的分子细节，并阐明 ctnnbl1调节拟靶活性的机制。 可以预见，这项研究的结果将与理解机制高度相关 抗体多样性的产生以及创建新的治疗策略以最小化的基础 基因组疾病和癌症。