CONTROL OF C-MYC TRANSCRIPTION IN AGGRESSIVE LYMPHOMAS

我建议从分子水平研究c-myc的激活机制。 T(8；14)在Burkitt淋巴瘤和转化性淋巴瘤中的水平 易位。我们的目标是更好地理解 B细胞恶性转化的机制。 A.转录调控分子机制的鉴定 C-myc基因在B细胞中的表达。 将使用体内足迹和体外蛋白质-DNA结合研究 寻找正常B细胞c-myc表达的重要调控区 不含t(8；14)易位的B细胞系。 结合在体内的蛋白质的特性将由一个 紫外光交联和免疫沉淀技术 杂交分析。将对这些地点进行功能评估 在B细胞系中进行瞬时转基因实验。 B.转录调控分子机制的鉴定 伯基特淋巴瘤的c-myc基因。条件得到了优化 从正常c-myc基因中分离易位基因 几个伯基特细胞系。转录结合的差异 已经发现了两个等位基因之间的因素，这些研究将是 继续。蛋白的鉴定和性质研究 体内的结合将按照A.甲基化状态的描述进行测定 将对每个等位基因进行比较。 C.转录调控分子机制的鉴定 C-myc和bcl2基因在转化性淋巴瘤及其低度恶性中的表达 对口单位。C-myc和bc1-2的解除调控 免疫球蛋白基因座的研究将如B。 D.调控基因中发现的突变对c-myc表达的影响 地区。任何突变对调节因子结合的影响 蛋白质将通过体内和体外研究进行研究。 E.免疫球蛋白基因座在激活c-myc表达中的作用。 复制c-myc易位的模型构建将是 构建并用于定义免疫球蛋白的重要区域 轨迹。 这项提案的成功完成将是第一个 例如，参与转录的分子机制 通过易位激活癌基因是已知的。抄写 将确定激活c-myc所需的因子。 以及免疫球蛋白基因座上负责这一点的区域 将会被定义。

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.