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.

摘要： 激活诱导脱氨酶(AID)是一种突变脊椎动物抗体基因的酶。这些 援助介导的突变是有机体产生识别和识别特定抗体所必需的 根除各种微生物感染。然而，如果艾滋病没有得到适当的监管，它可能会改变DNA 对正常细胞功能和致病很重要的基因序列。事实上，一些淋巴瘤和 白血病是由艾滋病的异常活动引起的。AID如何仅在抗体中改变DNA序列 人们对基因而不是其他基因知之甚少。最近，连接素样蛋白1(CTNNBL1)被 被发现与艾滋病有身体上的相互作用。CTNNBL1的生理功能知之甚少。然而， AID需要与CTNNBL1相互作用才能改变抗体基因中的DNA序列。 在这项研究中，高通量方法的组合包括下一代测序和 微阵列分析和有针对性的生化和免疫学分析将用于调查艾滋病是如何 特别是突变抗体基因，特别是哪些基因组区域可以获得AID，这两个基因都是 以及在没有CTNNBL1的情况下。这些实验还将检测到转录的任何变化 和由于缺乏CTNNBL1而引起的剪接，从而揭示了正常的细胞功能 CTNNBL1.将进行质谱分析、诱变分析和免疫学实验 确定AID与CTNNBL1相互作用的分子细节，并阐明 CTNNBL1调控AID靶向活性的机制 预计这项研究的结果将与理解这些机制高度相关。 在产生抗体多样性的基础上，并在创造新的治疗策略方面尽量减少 基因组疾病和癌症。