Function of the AID C terminus in Ig class switching

AID C 末端在 Ig 类别转换中的功能

基本信息

项目摘要

DESCRIPTION (provided by applicant): This is an application for an exploratory grant to build on our preliminary results suggesting that the C terminus of activation-induced cytidine deaminase (AID) is important for the recombination step during antibody class switch recombination (CSR). It has been known for several years that the C terminal 10 amino acids of AID are very important for CSR, although they do not appear to have any role during somatic hypermutation (SHM) of antibody genes, a process also dependent upon AID. Also, the AID C terminus is important for preventing chromosomal translocations between the IgH and c-myc loci. We have obtained novel results indicating that in splenic B cells induced to undergo CSR, AID binds to Ig switch (S) regions cooperatively with other enzymes involved in introducing DNA breaks into S regions, specifically UNG and Msh2-Msh6, and that this binding is dependent upon the AID C terminus. Using retroviral over-expression in aid-/- mouse splenic B cells of AID, we detect AID binding to S? and S?? in chromatin immunoprecipitation (ChIP) assays, whereas C terminal deleted AID (?AID) does not ChIP with S region DNA. Likewise, both UNG and Msh2-Msh6 are also detected by ChIP at S? in aid-/- cells expressing ?AID, but they are not detected in cells expressing AID, suggesting that the binding of these repair proteins depends on the AID C terminus, and the binding of AID and these proteins might be cooperative, i.e. co-dependent. Consistent with the hypothesis that these proteins bind cooperatively to S? DNA, in ung-/-aid-/- B cells or in msh2-/-aid-/- B cells, transduced full lengt AID cannot be detected at S? by ChIP. These results suggest that in order for AID and UNG and Msh2-Msh6 to bind sufficiently stably to be detected by ChIP at S regions, they must bind DNA cooperatively with each other, and this cooperative binding depends on the AID C terminus. We propose to test the hypotheses (1) that the AID C terminus recruits UNG and Msh2-Msh6 to S regions via an intermediary protein, and (2) that the C terminus of AID is important for recruiting UNG and Msh2-Msh6 to S regions during the G1 phase of the cell cycle, and (3) that the C terminus of AID is important for steps subsequent to formation of DNA breaks that direct CSR towards non-homologous end-joining (NHEJ). In order to test these hypotheses we propose 3 Specific Aims: 1) To investigate the mechanism of interaction between AID and UNG and Msh2-Msh6. 2) To determine if the recruitment of UNG and Msh2-Msh6 by AID is important for creating DSBs in S regions during G1 phase and also for their repair during G1 phase. In normal splenic B cells, AID-dependent S? DSBs are restricted to the G1 phase. However, in other cell types, UNG and Msh2- Msh6 are recruited by the DNA replication complex to DNA during S phase. 3) To determine if the AID C terminus is important for recruiting enzymes involved in NHEJ to S regions and thereby directing CSR toward NHEJ. PUBLIC HEALTH RELEVANCE: This project investigates the function of the enzyme (AID) that initiates antibody class switching, which is required for generation of an effective antibody immune response. This enzyme initiates formation of breaks in DNA which when everything goes well leads to an effective immune response. However, AID can cause collateral damage, which leads to chromosomal deletions and translocations and to B cell lymphomas, the most common type of cancer. We will investigate how AID is regulated to introduce DNA breaks that lead to proper antibody class switching.
描述(由申请人提供):这是一项探索性拨款的申请,以我们的初步结果为基础,该结果表明,激活诱导的胞苷脱氨酶(AID)的C末端对于抗体类别转换重组(CSR)中的重组步骤是重要的。已知AID的C端10个氨基酸对CSR非常重要,尽管它们在抗体基因的体细胞超突变(SHM)过程中似乎没有任何作用,这一过程也依赖于AID。此外,AID C末端对于防止IgH和c-myc基因座之间的染色体易位也很重要。我们的新结果表明,在被诱导为CSR的脾B细胞中,AID与参与将DNA断裂引入S区域的其他酶协同结合到Ig开关(S)区域,特别是UNG和Msh2-MSH6,并且这种结合依赖于AID C末端。利用逆转录病毒在艾滋病小鼠脾B细胞中的过度表达,检测艾滋病病毒与S的结合。S呢??在染色质免疫沉淀(CHIP)检测中,C端缺失的AID(?AID)不与S区域DNA发生芯片反应。同样,在S的芯片中也检测到了UNG和Msh2-MSH6。但在表达AID的细胞中未检测到,提示这些修复蛋白的结合依赖于AID的C末端,AID与这些蛋白的结合可能是协同的,即相互依赖的。与这些蛋白质与S协同结合的假设一致吗?S未检测到转导全长AID的Un-/-Aid-/-B细胞或MsH2-/-Aid-/-B细胞中的DNA?用芯片。这些结果表明,Aid、UNG和Msh2-MSH6要足够稳定地结合到S区域的芯片上,它们必须相互协同结合DNA,这种协同结合依赖于AID C末端。我们建议检验假设(1)AID的C末端通过中间蛋白将UNG和Msh2-MSH6招募到S区域,(2)在细胞周期的G1期,AID的C末端对于将UNG和Msh2-MSH6招募到S区域是重要的,(3)AID的C末端对于DNA断裂的形成是重要的,DNA断裂将CSR导向非同源末端连接(NHEJ)。为了验证这些假说,我们提出了三个具体目标:1)研究AID与UNG和Msh2-MSH6的相互作用机制。2)研究AID对UNG和Msh2-MSH6的募集是否对S区G1期双链断裂的产生和G1期双链断裂的修复起重要作用。在正常脾B细胞中,艾滋病依赖的S?双链断裂被限制在G1相。然而,在其他类型的细胞中,UNG和Msh2-MSH6在S期被DNA复制复合体招募到DNA中。3)确定AID C末端是否对将NHEJ相关酶招募到S区域,从而将CSR导向NHEJ起重要作用。 公共卫生相关性:该项目调查启动抗体类别转换的酶(AID)的功能,这是产生有效的抗体免疫反应所必需的。这种酶启动DNA断裂的形成,当一切顺利时,会产生有效的免疫反应。然而,艾滋病会造成附带损害,导致染色体缺失和易位,并导致最常见的癌症--B细胞淋巴瘤。我们将研究AID如何被调控以引入DNA断裂,从而导致适当的抗体类别转换。

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)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Janet M. Stavnezer其他文献

Janet M. Stavnezer的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Janet M. Stavnezer', 18)}}的其他基金

Molecular Basis of Immunoglobulin Heavy Chain Switch
免疫球蛋白重链开关的分子基础
  • 批准号:
    8090512
  • 财政年份:
    2010
  • 资助金额:
    $ 20.76万
  • 项目类别:
c-myc DNA breaks and c-myc-IgH locus translocations: roles of AID and oxidation
c-myc DNA 断裂和 c-myc-IgH 基因座易位:AID 和氧化的作用
  • 批准号:
    7865093
  • 财政年份:
    2010
  • 资助金额:
    $ 20.76万
  • 项目类别:
c-myc DNA breaks and c-myc-IgH locus translocations: roles of AID and oxidation
c-myc DNA 断裂和 c-myc-IgH 基因座易位:AID 和氧化的作用
  • 批准号:
    8097530
  • 财政年份:
    2010
  • 资助金额:
    $ 20.76万
  • 项目类别:
Molecular Basis of Immunoglobulin Heavy Chain Switch
免疫球蛋白重链开关的分子基础
  • 批准号:
    7846563
  • 财政年份:
    2009
  • 资助金额:
    $ 20.76万
  • 项目类别:
Isotype specific regulation of lg class switching
LG 类别转换的同种型特异性调节
  • 批准号:
    7140383
  • 财政年份:
    2005
  • 资助金额:
    $ 20.76万
  • 项目类别:
Isotype specific regulation of lg class switching
LG 类别转换的同种型特异性调节
  • 批准号:
    6965565
  • 财政年份:
    2005
  • 资助金额:
    $ 20.76万
  • 项目类别:
DNA repair and lg class switching
DNA 修复和 LG 类别转换
  • 批准号:
    7012289
  • 财政年份:
    2005
  • 资助金额:
    $ 20.76万
  • 项目类别:
DNA repair and lg class switching
DNA 修复和 LG 类别转换
  • 批准号:
    7172597
  • 财政年份:
    2005
  • 资助金额:
    $ 20.76万
  • 项目类别:
DNA repair and lg class switching
DNA 修复和 LG 类别转换
  • 批准号:
    6853179
  • 财政年份:
    2005
  • 资助金额:
    $ 20.76万
  • 项目类别:
INDUCTION OF IG C EPSILON & C GAMMA 1 BY IL4 & CD40L
IG C Epsilon 感应
  • 批准号:
    6510760
  • 财政年份:
    1998
  • 资助金额:
    $ 20.76万
  • 项目类别:

相似海外基金

Double Incorporation of Non-Canonical Amino Acids in an Animal and its Application for Precise and Independent Optical Control of Two Target Genes
动物体内非规范氨基酸的双重掺入及其在两个靶基因精确独立光学控制中的应用
  • 批准号:
    BB/Y006380/1
  • 财政年份:
    2024
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Research Grant
Quantifying L-amino acids in Ryugu to constrain the source of L-amino acids in life on Earth
量化 Ryugu 中的 L-氨基酸以限制地球生命中 L-氨基酸的来源
  • 批准号:
    24K17112
  • 财政年份:
    2024
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Molecular recognition and enantioselective reaction of amino acids
氨基酸的分子识别和对映选择性反应
  • 批准号:
    23K04668
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Basic research toward therapeutic strategies for stress-induced chronic pain with non-natural amino acids
非天然氨基酸治疗应激性慢性疼痛策略的基础研究
  • 批准号:
    23K06918
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Molecular mechanisms how arrestins that modulate localization of glucose transporters are phosphorylated in response to amino acids
调节葡萄糖转运蛋白定位的抑制蛋白如何响应氨基酸而被磷酸化的分子机制
  • 批准号:
    23K05758
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Design and Synthesis of Fluorescent Amino Acids: Novel Tools for Biological Imaging
荧光氨基酸的设计与合成:生物成像的新工具
  • 批准号:
    2888395
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Studentship
Collaborative Research: RUI: Elucidating Design Rules for non-NRPS Incorporation of Amino Acids on Polyketide Scaffolds
合作研究:RUI:阐明聚酮化合物支架上非 NRPS 氨基酸掺入的设计规则
  • 批准号:
    2300890
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
    Continuing Grant
Structurally engineered N-acyl amino acids for the treatment of NASH
用于治疗 NASH 的结构工程 N-酰基氨基酸
  • 批准号:
    10761044
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
Lifestyle, branched-chain amino acids, and cardiovascular risk factors: a randomized trial
生活方式、支链氨基酸和心血管危险因素:一项随机试验
  • 批准号:
    10728925
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
Single-molecule protein sequencing by barcoding of N-terminal amino acids
通过 N 端氨基酸条形码进行单分子蛋白质测序
  • 批准号:
    10757309
  • 财政年份:
    2023
  • 资助金额:
    $ 20.76万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了