CONTROL OF C MYC TRANSCRIPTION IN HIGH GRADE LYMPHOMAS

高级淋巴瘤中 C MYC 转录的控制

• 批准号: 2654206
• 负责人: LINDA M BOXER
• 金额: $ 18.44万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-15 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2654206
• 关键词: B lymphocyte; Burkitt's lymphoma; DNA binding protein; DNA footprinting; DNA methylation; chemical binding; chromosome translocation; crosslink; gene expression; genetic models; genetic transcription; human tissue; immunoglobulin genes; immunoprecipitation; lymphoma; neoplasm /cancer genetics; neoplastic transformation; nucleic acid hybridization; oncogenes; protooncogene; tissue /cell culture; transcription factor; transfection

DESCRIPTION: (adapted from the investigator's abstract) The applicant proposes to study the mechanism of activation of c-myc at a molecular level in both Burkitt's lymphoma and transformed lymphomas with the t(8;14) translocation. The goal is to reach a better understanding of the mechanisms of malignant transformation of B cells. A. Identification of the molecular mechanisms of transcriptional control of the c-myc gene in B cells. In vivo footprinting and in vitro protein-DNA binding studies will be used to locate regulatory regions important for c-myc expression in normal B cells and in B cell lines that do not contain the t(8;14) translocation. The identity of the proteins that bind in vivo will be determined by a technique of UV crosslinking and immunoprecipitation followed by hybridization analysis. Functional assessment of the sites will be performed with transient transfection experiments in B cell lines. B. Identification of the molecular mechanisms of transcriptional control of the c-myc gene in Burkitt's lymphomas. Conditions have been optimized for the separation of the translocation from the normal c-myc gene in several Burkitt's cell lines. Differences in the binding of transcription factors between the two alleles have been found and these studies will be continued. The identification and characterization of the proteins that bind in vivo will be determined as described in A. The methylation status of each allele will be compared. C. Identification of the molecular mechanisms of transcriptional control of the c-myc and bcl-2 genes in transformed lymphomas and their low grade counterparts. The deregulation of both c-myc and bcl-2 by the immunoglobulin locus will be studied as outlined in B. D. Effect on c-myc expression of mutations identified in the regulatory regions. The effects of any mutations on the binding of regulatory proteins will be investigated by in vivo and in vitro studies. E. Role of the immunoglobulin locus in the activation of c-myc expression. Model constructs which reproduce the c-myc translocations will be constructed and used to define the important regions of the immunoglobulin locus. The successful completion of this proposal will represent the first instance where the molecular mechanisms involved in the transcriptional activation of an oncogene by translocation are known. The transcription factors that are required for the activation of c-myc will be identified and the regions of the immunoglobulin locus which are responsible for this will be defined.

描述：（根据调查员的摘要改编）申请人 提议研究分子上C-MYC激活的机制 伯基特淋巴瘤和转化淋巴瘤的水平 t（8; 14）易位。目标是更好地理解 B细胞恶性转化的机制。 A.鉴定转录控制的分子机制 B细胞中的C-Myc基因。体内足迹和体外 蛋白-DNA结合研究将用于定位调节区域 对于正常B细胞​​和B细胞系中的C-MYC表达很重要 不包含t（8; 14）易位。的身份 结合体内结合的蛋白质将由紫外线技术确定 交联和免疫沉淀，然后进行杂交分析。 将对站点进行功能评估 在B细胞系中的转染实验。 B.鉴定转录控制的分子机制 Burkitt淋巴瘤中的C-Myc基因。条件已经存在 优化了从正常C-MYC分离易位的 基因在几个伯基特的细胞系中。结合的差异 已经发现了两个等位基因之间的转录因子， 研究将继续。识别和表征 结合体内结合的蛋白将如图所述确定。 将比较每个等位基因的甲基化状态。 C.鉴定转录控制的分子机制 转化的淋巴瘤中的C-Myc和Bcl-2基因及其低 等级对应。通过 免疫球蛋白基因座将如B中概述。 D.对调节性突变的C-MYC表达的影响 地区。任何突变对调节结合的影响 蛋白质将通过体内和体外研究研究。 E.免疫球蛋白基因座在C-MYC激活中的作用 表达。重现C-MYC易位的模型构造 将被构造并用来定义重要区域 免疫球蛋白基因座。 该提案的成功完成将代表第一个 实例分子机制涉及转录 已知通过易位激活致癌基因。转录 将确定激活C-MYC所需的因素 以及负责的免疫球蛋白基因座的区域 这将被定义。