CONTROL OF C-MYC TRANSCRIPTION IN AGGRESSIVE LYMPHOMAS

侵袭性淋巴瘤中 C-MYC 转录的控制

基本信息

批准号：
6236697
负责人：
LINDA M BOXER
金额：
$ 25.1万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-06-27 至 1998-03-31
项目状态：
已结题

项目摘要

I propose 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 th 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 translocated 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 ont he 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.

我建议从分子水平上研究c-myc的激活机制伯基特淋巴瘤和转化淋巴瘤中 t(8;14) 的水平易位。目标是更好地了解 B细胞恶性转化的机制。 A. 转录控制分子机制的鉴定 B 细胞中的 c-myc 基因。将使用体内足迹和体外蛋白质-DNA 结合研究定位对正常 B 中 c-myc 表达重要的调节区域细胞和不包含 t(8;14) 易位的 B 细胞系。体内结合的蛋白质的身份将由紫外交联和免疫沉淀技术，然后杂交分析。场地的功能评估将在 B 细胞系中进行瞬时转染实验。 B. 转录控制分子机制的鉴定伯基特淋巴瘤中的 c-myc 基因。条件已优化用于从正常 c-myc 基因中分离转位几种伯基特细胞系。转录结合的差异两个等位基因之间的因素已经被发现，这些研究将被持续。蛋白质的鉴定和表征体内结合将按照 A 中的描述进行测定。甲基化状态将比较每个等位基因的。 C. 转录控制分子机制的鉴定转化淋巴瘤中的 c-myc 和 bcl-2 基因及其低级别同行。 c-myc 和 bcl-2 的放松管制将按照 B 中概述的方式研究免疫球蛋白位点。 D. 监管中发现的突变对 c-myc 表达的影响地区。任何突变对监管结合的影响将通过体内和体外研究来研究蛋白质。 E. 免疫球蛋白基因座在 c-myc 表达激活中的作用。再现 c-myc 易位的模型构建体将是构建并用于定义免疫球蛋白的重要区域轨迹。该提案的成功完成将成为第一个例如，涉及转录的分子机制通过易位激活癌基因是已知的。转录将确定激活 c-myc 所需的因子以及负责此的免疫球蛋白基因座区域将被定义。