Mechanism and regulation of DNA end processing in V(D)J recombination and repair

V(D)J重组和修复中DNA末端加工的机制和调控

• 批准号: 8488394
• 负责人: JoAnn Sekiguchi
• 金额: $ 32.47万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8488394
• 关键词: Alleles; Antigen Receptors; Antigens; Arts; B-Lymphocytes; Binding; Biological Models; Catalytic Domain; Cells; Chromosomal translocation; Code; Codon Nucleotides; Complex; DNA; DNA Double Strand Break; DNA Repair; DNA-dependent protein kinase; Defect; Deoxyribonucleases; Development; Disease; Double Strand Break Repair; Event; Gene Rearrangement; Genes; Genetic; Genome Stability; Genomic Instability; Goals; Human; Immune system; Immunodeficiency and Cancer; Immunologic Deficiency Syndromes; Individual; Inherited; Investigation; J segment gene; Knock-in Mouse; Lead; Ligation; Lymphocyte; Lymphoid; Maintenance; Malignant lymphoid neoplasm; Mediating; Molecular; Mus; Mutation; Nonhomologous DNA End Joining; Nonsense Codon; Nonsense Mutation; Oncogenic; Patients; Pharmaceutical Preparations; Phenotype; Play; Predisposition; Process; Radiation Tolerance; Reading; Receptor Gene; Regulation; Role; ST5 gene; Severe Combined Immunodeficiency; Severities; Syndrome; System; Systems Development; T-Lymphocyte; Testing; Therapeutic; Translations; V(D)J Recombination; artemis; congenital immunodeficiency; development of lymphoid malignancy; effective therapy; endonuclease; human disease; in vivo; insight; mouse model; mutant; novel; novel therapeutics; nuclease; premature; public health relevance; receptor; recombinational repair; repaired; therapeutic effectiveness; tumor; tumorigenesis; variable region gene

DESCRIPTION (provided by applicant): B and T lymphocytes, the primary cells of the adaptive immune system, provide a major line of defense against a myriad of foreign molecules by virtue of antigen-specific receptors. The vast diversity amongst the antigen receptors genes is generated through V(D)J recombination, a process in which individual V, D, and J gene segments are rearranged. Defects in V(D)J recombination lead to combined immunodeficiencies in human patients and can also result in lymphoid malignancies. One factor that plays a critical role during V(D)J recombination is the non-homologous end joining (NHEJ) DNA repair factor, Artemis. Artemis was initially discovered as the gene inactivated in a human radiosensitive severe combined immunodeficiency syndrome. Hypomorphic Artemis alleles have also been identified in patients and are associated with combined immunodeficiencies of varying severity. Artemis plays a vital role as a DNA nuclease that processes broken ends during V(D)J recombination as well as general double strand break repair prior to ligation. Its intrinsic exo- and endonucleolytic activities are modulated by interaction with the DNA-dependent protein kinase catalytic subunit NHEJ factor. However, the precise mechanisms by which Artemis activities are regulated in vivo are not well understood. The major goals of this proposal are to gain a better understanding of the mechanisms and regulation of DNA end processing during V(D)J rearrangements and general DNA repair. Three specific aims are proposed. Aim 1 is to define the molecular mechanisms involved in activating and regulating Artemis endonucleolytic activities. Aim 2 is to elucidate the molecular mechanisms underlying tumorigenesis caused by a hypomorphic, Artemis disease allele, P70, using the mouse as a model system. This premature translation termination mutation leads to partial immunodeficiency and predisposition to lymphoid malignancy in human patients. In addition, the genetic interactions between Artemis, ATM and Mre11 will be examined. Aim 3 will capitalize on the previously developed Artemis-P70 mouse model, which provides a valuable in vivo system to define the impact of therapeutics that allow read-through of nonsense mutations. The impact of this emerging class of drugs on the DNA repair, genome instability, V(D)J recombination, and lymphocyte development phenotypes in mutant cells and mice will be determined. Together, these studies will provide important insights into the molecular events that occur during V(D)J coding end joining and general DSB repair, elucidate the impact of aberrant V(D)J end processing on lymphocyte development and tumor predisposition, and potentially identify novel therapeutics for primary immunodeficiencies caused by nonsense mutations.

描述（由申请人提供）：B 和 T 淋巴细胞是适应性免疫系统的主要细胞，通过抗原特异性受体提供针对无数外来分子的主要防线。抗原受体基因之间的巨大多样性是通过 V(D)J 重组产生的，在该过程中，各个 V、D 和 J 基因片段重新排列。 V(D)J 重组缺陷会导致人类患者出现联合免疫缺陷，也可能导致淋巴恶性肿瘤。在 V(D)J 重组过程中起关键作用的因素之一是非同源末端连接 (NHEJ) DNA 修复因子 Artemis。 Artemis 最初被发现是在人类放射敏感性严重联合免疫缺陷综合征中失活的基因。亚形阿耳忒弥斯等位基因也在患者中被发现，并与不同严重程度的联合免疫缺陷相关。 Artemis 作为 DNA 核酸酶发挥着至关重要的作用，可在 V(D)J 重组过程中处理断裂末端，以及在连接前进行一般双链断裂修复。其内在的核酸外切和核酸内切活性通过与 DNA 依赖性蛋白激酶催化亚基 NHEJ 因子的相互作用进行调节。然而，Artemis 活性在体内调节的精确机制尚不清楚。 该提案的主要目标是更好地了解 V(D)J 重排和一般 DNA 修复过程中 DNA 末端加工的机制和调节。提出了三个具体目标。目标 1 是确定参与激活和调节 Artemis 核酸内切活性的分子机制。目标 2 是使用小鼠作为模型系统，阐明由亚形态性 Artemis 疾病等位基因 P70 引起的肿瘤发生的分子机制。这种过早的翻译终止突变会导致人类患者部分免疫缺陷和淋巴恶性肿瘤的易感性。此外，Artemis、ATM 和 Mre11 之间的遗传相互作用也将得到检查。 Aim 3 将利用之前开发的 Artemis-P70 小鼠模型，该模型提供了一个有价值的体内系统来定义允许读取无义突变的疗法的影响。这类新兴药物对突变细胞和小鼠的 DNA 修复、基因组不稳定性、V(D)J 重组和淋巴细胞发育表型的影响将得到确定。总之，这些研究将为 V(D)J 编码末端连接和一般 DSB 修复过程中发生的分子事件提供重要见解，阐明异常 V(D)J 末端加工对淋巴细胞发育和肿瘤易感性的影响，并可能确定针对无义突变引起的原发性免疫缺陷的新疗法。

项目成果

A hypomorphic Artemis human disease allele causes aberrant chromosomal rearrangements and tumorigenesis.

亚型阿耳忒弥斯人类疾病等位基因会导致异常的染色体重排和肿瘤发生。

• DOI: 10.1093/hmg/ddq524
• 发表时间: 2011-02-15
• 期刊: Human molecular genetics
• 影响因子: 3.5
• 作者: Cheryl Jacobs;Ying Huang;T. Masud;William Lu;Gerwin H. Westfield;W. Giblin;J. Sekiguchi
• 通讯作者: J. Sekiguchi

Impact of a hypomorphic Artemis disease allele on lymphocyte development, DNA end processing, and genome stability.

亚型阿尔忒弥斯病等位基因对淋巴细胞发育、DNA 末端加工和基因组稳定性的影响。

• 发表时间: 2009-04-13
• 作者: Huang, Ying;Giblin, William;Kubec, Martina;Westfield, Gerwin;St Charles, Jordan;Chadde, Laurel;Kraftson, Stephanie;Sekiguchi, JoAnn
• 通讯作者: Sekiguchi, JoAnn

The SNM1B/APOLLO DNA nuclease functions in resolution of replication stress and maintenance of common fragile site stability.

SNM1B/APOLLO DNA 核酸酶的作用是解决复制压力和维持常见脆弱位点的稳定性。

• DOI: 10.1093/hmg/ddt340
• 发表时间: 2013-12-15
• 期刊: Human molecular genetics
• 影响因子: 3.5
• 作者: Jennifer M O Mason;I. Das;M. Arlt;N. Patel;Stephanie J Kraftson;T. Glover;J. Sekiguchi
• 通讯作者: J. Sekiguchi

Structural and functional interaction between the human DNA repair proteins DNA ligase IV and XRCC4.

人类 DNA 修复蛋白 DNA 连接酶 IV 和 XRCC4 之间的结构和功能相互作用。

• 发表时间: 2009-06
• 影响因子: 5.3
• 作者: Wu, Peï;Frit, Philippe;Meesala, SriLakshmi;Dauvillier, Stéphanie;Modesti, Mauro;Andres, Sara N;Huang, Ying;Sekiguchi, JoAnn;Calsou, Patrick;Salles, Bernard;Junop, Murray S
• 通讯作者: Junop, Murray S

Expansion of immunoglobulin-secreting cells and defects in B cell tolerance in Rag-dependent immunodeficiency.

Rag 依赖性免疫缺陷中免疫球蛋白分泌细胞的扩增和 B 细胞耐受性缺陷。

• 发表时间: 2010-07-05
• 作者: Walter, Jolan E;Rucci, Francesca;Patrizi, Laura;Recher, Mike;Regenass, Stephan;Paganini, Tiziana;Keszei, Marton;Pessach, Itai;Lang, Philipp A;Poliani, Pietro Luigi;Giliani, Silvia;Al;Cowan, Morton J;Puck, Jennifer M;Bleesing, Ja
• 通讯作者: Bleesing, Ja