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: 8591624
• 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/8591624
• 关键词: Address; Antibodies; Antibody Affinity; Antibody Diversity; Antigens; B-Cell Lymphomas; B-Lymphocytes; Biochemical; Cancer Etiology; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromosomal translocation; 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; activation-induced cytidine deaminase; cis acting element; immune function; insight; leukemia/lymphoma; microbial; mutant; next generation sequencing; novel; novel therapeutics; pathogen; protein activation; research study

DESCRIPTION (provided by applicant): 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 2 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.

描述（由申请人提供）：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是生物体生成特定抗体以识别和消除大量微生物感染的特定抗体所必需的。但是，如果没有适当调节辅助，它可以改变对正常细胞功能重要的基因的DNA序列并引起疾病。实际上，某些淋巴瘤和白血病是由异常活动引起的。辅助如何仅在抗体基因中更改DNA序列，而在其他基因中不能改变DNA序列。最近，发现Catenin 2像1蛋白（CTNNBL1）与AID物理相互作用。 CTNNBL1的生理功能闻名很少。然而，与CTNNBL1相互作用是需要在抗体基因中更改DNA序列所必需的。在这项研究中，将使用高吞吐量方法的结合，包括下一代测序和微阵列分析以及靶向的生化和免疫学分析，用于研究如何在存在和不存在CTNNBL1的情况下特异性突变的抗体基因，尤其是在存在和缺乏CTNNBL1的情况下可以访问的基因组区域，尤其是基因组区域。这些实验还将检测到由CTNNBL1缺乏引起的转录和剪接的任何改变，从而在CTNNBL1的正常细胞功能上脱光灯。将进行质谱，诱变分析和免疫学实验，以确定AID与CTNNBL1之间相互作用的分子细节，并阐明CTNNBL1调节靶向AID活动的机制。可以预料，这项研究的结果将与理解抗体多样性产生的机制以及创建新的治疗策略以最大程度地减少基因组疾病和癌症的机制高度相关。

项目成果

Intrinsic transcriptional heterogeneity in B cells controls early class switching to IgE.

• DOI: 10.1084/jem.20161056
• 发表时间: 2017-01
• 期刊: The Journal of experimental medicine
• 作者: Wu YL;Stubbington MJ;Daly M;Teichmann SA;Rada C
• 通讯作者: Rada C

DNA deaminases induce break-associated mutation showers with implication of APOBEC3B and 3A in breast cancer kataegis.

• DOI: 10.7554/elife.00534
• 发表时间: 2013-04-16
• 期刊: eLife
• 影响因子: 7.7
• 作者: Taylor BJ;Nik-Zainal S;Wu YL;Stebbings LA;Raine K;Campbell PJ;Rada C;Stratton MR;Neuberger MS
• 通讯作者: Neuberger MS