Molecular Mechanisms of Class Switch Recombination
类别转换重组的分子机制
基本信息
- 批准号:8386894
- 负责人:
- 金额:$ 40.08万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2008
- 资助国家:美国
- 起止时间:2008-12-15 至 2014-02-28
- 项目状态:已结题
- 来源:
- 关键词:AddressAlanineAllergic DiseaseAntibodiesAntibody FormationAutoimmune DiseasesB lymphoid malignancyB-LymphocytesBiochemical GeneticsBiological AssayCancer EtiologyCellsChromosomal translocationChromosome PairingChromosomesComplexCytidine DeaminaseCytogeneticsDNADNA Double Strand BreakDNA RepairDNA SequenceDNA StructureDeaminationDiseaseDouble Strand Break RepairExonsExtrinsic asthmaFrequenciesG22P1 geneGene RearrangementGenesGeneticGenetic ModelsGenetic TranscriptionHeavy-Chain ImmunoglobulinsHigher Order Chromatin StructureIGH@ gene clusterIgEImmune systemImmunoglobulin Class SwitchingImmunoglobulin Joining RegionImmunoglobulin Somatic HypermutationImmunoglobulin Switch RecombinationImmunoglobulin Variable RegionImmunologic Deficiency SyndromesImmunologyIndiumLinkLymphomaMalignant NeoplasmsMediatingMethodsModificationMolecularMolecular ConformationMusMutant Strains MiceMutationNonhomologous DNA End JoiningOncogenesOutcomePathogenesisPathway interactionsPhosphorylationProcessProductionProteinsReagentRegulationReporterRoleSeriesSerineSingle-Stranded DNASiteSpecificityTestingTransgenic OrganismsVaccinesWorkYeastsZebrafishactivation-induced cytidine deaminasechromatin immunoprecipitationds-DNAendonucleasein vitro Assayin vivoinsightmutantnovelnovel strategiesprotein complexrepairedreplication factor Aresponse
项目摘要
This application proposes studies of the mechanisms of immunoglobulin heavy chain (IgH) class switch
recombination (CSR) and Somatic Hypermutation (SHM). We have shown that Activation Induced Cytidine
Deaminase (AID), the initiator of CSR, is a single strand DNA (ssDNA) specific cytidine deaminase and we
employed a series of novel biochemical and genetic approaches to elucidate mechanisms by which AID gains
access to transcribed double strand (ds)DNA sequences in the context transcription-generated ssDNA
structures and/or certain AID modifications or co-factors. We also showed that CSR may employ general
processes for synapsis of AID-initiated DNA double strand breaks (DSBs), that general DNA repair factors
function in CSR, and that two distinct end-joining pathways fuse S region breaks to complete CSR. Our
current proposal builds on these observations in the context of three specific Aims. Our first aim proposes use
of biochemical and genetic approaches to elucidate basic mechanisms of AID function and regulation. In this
regard, we developed methods to purify AID from normal B cells, in vitro assays for transcription-dependent
AID deamination of dsDNA DNA, and genetic approaches to evaluate in vivo AID functions elucidated
biochemically. Our second aim addresses mechanisms by which DNA sequences influence AID activity and
its outcome. For these studies, we developed targeted mutation assays to replace endogenous IgH class
switch (S) regions and exons encoding IgH variable regions with test sequences that will allow us to determine
how substrate sequences influence activities of AID and other relevant factors in CSR and SHM. Together, the
complementary biochemical and genetic assays of Aims 1 and 2 offer a powerful approach for elucidating
factors and mechanisms involved in initiation and regulation of IgH CSR and SHM. A third proposed aim is to
elucidate processes involved in the repair of AID induced DSBs to complete CSR. For these studies, we again
have developed a large array of reagents and novel approaches, including cytogenetic methods to follow CSR
related breaks in chromosomes, novel genetic approaches to study factors involved in long range synapsis of
DSBs, and genetic models to elucidate DSB repair pathways that complete IgH CSR. Our proposed studies
should provide novel insights into the mechanism of antibody production via IgH CSR and, therefore, be
relevant to understanding immunodeficiencies, vaccine immunology, and autoimmune diseases. As CSR is
required for IgE production, the work will also be relevant to understanding pathogenesis of allergic diseases
and asthma. Finally, the work is relevant to B cell malignancies as they often involve chromosomal
translocations that link translocated oncogenes to IgH S regions via aberrant CSR.
本申请提出了免疫球蛋白重链 (IgH) 类别转换机制的研究
重组(CSR)和体细胞超突变(SHM)。我们已经证明激活诱导胞苷
脱氨酶 (AID) 是 CSR 的引发剂,是一种单链 DNA (ssDNA) 特异性胞苷脱氨酶,我们
采用了一系列新颖的生化和遗传学方法来阐明 AID 获益的机制
在转录生成的 ssDNA 中获取转录的双链 (ds)DNA 序列
结构和/或某些 AID 修饰或辅因子。我们还表明,企业社会责任可以采用一般
AID 引发的 DNA 双链断裂 (DSB) 的突触过程,即一般 DNA 修复因子
在 CSR 中发挥作用,并且两个不同的末端连接途径融合 S 区断裂以完成 CSR。我们的
当前的提案以三个具体目标的背景下的这些观察为基础。我们的首要目标是建议使用
生物化学和遗传学方法来阐明 AID 功能和调节的基本机制。在这个
为此,我们开发了从正常 B 细胞中纯化 AID 的方法,并进行转录依赖性体外测定
阐明双链 DNA 的 AID 脱氨基作用以及评估体内 AID 功能的遗传方法
生物化学上。我们的第二个目标是解决 DNA 序列影响 AID 活性的机制
它的结果。对于这些研究,我们开发了靶向突变检测来替代内源性 IgH 类
使用测试序列转换编码 IgH 可变区的 (S) 区域和外显子,这将使我们能够确定
底物序列如何影响 AID 的活性以及 CSR 和 SHM 中的其他相关因素。在一起,
目标 1 和目标 2 的互补生化和基因检测提供了一种强有力的方法来阐明
参与 IgH CSR 和 SHM 启动和调节的因素和机制。拟议的第三个目标是
阐明参与 AID 诱导 DSB 修复以完成 CSR 的过程。对于这些研究,我们再次
开发了大量试剂和新方法,包括遵循 CSR 的细胞遗传学方法
染色体的相关断裂,研究参与长程突触的因素的新遗传学方法
DSB 和遗传模型阐明完成 IgH CSR 的 DSB 修复途径。我们提出的研究
应该为通过 IgH CSR 产生抗体的机制提供新的见解,因此,
与了解免疫缺陷、疫苗免疫学和自身免疫性疾病相关。正如企业社会责任是
IgE 产生所需,这项工作也将与了解过敏性疾病的发病机制相关
和哮喘。最后,这项工作与 B 细胞恶性肿瘤相关,因为它们通常涉及染色体
通过异常 CSR 将易位癌基因与 IgH S 区域连接起来的易位。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Frederick W. Alt其他文献
Case of mistaken identity
认错人的情况
- DOI:
10.1038/428029a - 发表时间:
2004-03-04 - 期刊:
- 影响因子:48.500
- 作者:
Craig H. Bassing;Frederick W. Alt - 通讯作者:
Frederick W. Alt
Activated Ras signals differentiation and expansion of CD4+8+ thymocytes.
激活的 Ras 发出 CD4 8 胸腺细胞分化和扩增的信号。
- DOI:
10.1073/pnas.93.10.4683 - 发表时间:
1996 - 期刊:
- 影响因子:11.1
- 作者:
Wojciech Swat;Yoichi Shinkai;Hwei;L. Davidson;Frederick W. Alt - 通讯作者:
Frederick W. Alt
From the Cover: Abnormal development of Purkinje cells and lymphocytes in Atm mutant mice
封面图片:Atm 突变小鼠浦肯野细胞和淋巴细胞发育异常
- DOI:
- 发表时间:
2000 - 期刊:
- 影响因子:0
- 作者:
P. Borghesani;Frederick W. Alt;Andrea Bottaro;L. Davidson;Saime Aksoy;Gary Rathbun;Thomas M. Roberts;Wojciech Swat;R. Segal;Yansong Gu - 通讯作者:
Yansong Gu
RNA editing meets DNA shuffling
RNA 编辑与 DNA 改组相遇
- DOI:
10.1038/35024189 - 发表时间:
2000-09-07 - 期刊:
- 影响因子:48.500
- 作者:
Ming Tian;Frederick W. Alt - 通讯作者:
Frederick W. Alt
Sirt3 Regulates Fatty Acid Oxidation via Reversible Enzyme Deacetylation Hhs Public Access Supplementary Material
Sirt3 通过可逆酶脱乙酰作用调节脂肪酸氧化 Hhs 公共访问补充材料
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
M. Hirschey;Tadahiro Shimazu;E. Goetzman;Enxuan Jing;B. Schwer;David B. Lombard;Carrie A. Grueter;Charles Harris;S. Biddinger;O. Ilkayeva;Robert D. Stevens;Yu Li;A. Saha;N. Ruderman;J. Bain;C. Newgard;R. V. Farese;Frederick W. Alt;C. R. Kahn;E. Verdin - 通讯作者:
E. Verdin
Frederick W. Alt的其他文献
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{{ truncateString('Frederick W. Alt', 18)}}的其他基金
Role of DNA Double Strand Break Response in Suppression of Thymic Lymphoma
DNA 双链断裂反应在抑制胸腺淋巴瘤中的作用
- 批准号:
7780950 - 财政年份:2010
- 资助金额:
$ 40.08万 - 项目类别:
Mouse models of severe combined immunodeficiencies
严重联合免疫缺陷小鼠模型
- 批准号:
7614101 - 财政年份:2009
- 资助金额:
$ 40.08万 - 项目类别:
Mechanisms that Regulate Antibody Class Switch Recombination and Somatic Hypermutation
调节抗体类别转换重组和体细胞超突变的机制
- 批准号:
10392890 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
Molecular Mechanisms of Class Switch Recombination
类别转换重组的分子机制
- 批准号:
7743798 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
AID Targeting Mechanisms for IgH Switch Recombination and Somatic Hypermutation
IgH 开关重组和体细胞超突变的 AID 靶向机制
- 批准号:
9228317 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
Mechanisms that Regulate Antibody Class Switch Recombination and Somatic Hypermutation
调节抗体类别转换重组和体细胞超突变的机制
- 批准号:
10612752 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
Molecular Mechanisms of Class Switch Recombination
类别转换重组的分子机制
- 批准号:
7577240 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
Molecular Mechanisms of Class Switch Recombination
类别转换重组的分子机制
- 批准号:
8197214 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
AID Targeting Mechanisms for IgH Switch Recombination and Somatic Hypermutation
IgH 开关重组和体细胞超突变的 AID 靶向机制
- 批准号:
8697880 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
Molecular Mechanisms of Class Switch Recombination
类别转换重组的分子机制
- 批准号:
7995253 - 财政年份:2008
- 资助金额:
$ 40.08万 - 项目类别:
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