CONTROL OF C MYC TRANSCRIPTION IN HIGH GRADE LYMPHOMAS

描述：(改编自调查人员摘要)申请人 建议从分子水平研究c-myc的激活机制。 Burkitt淋巴瘤和转化性淋巴瘤中 T(8；14)易位。我们的目标是更好地理解 B细胞恶变的机制。 A.转录调控分子机制的鉴定 C-myc基因在B细胞中的表达。体内足迹和体外足迹 蛋白质-DNA结合研究将用于定位调控区域 C-myc在正常B细胞和B细胞系中的重要表达 不含t(8；14)易位的基因。这些人的身份 体内结合的蛋白质将通过紫外光技术确定 交联法和免疫沉淀法，然后杂交分析。 将对现场进行功能评估，包括临时 在B细胞系中进行了基因转导实验。 B.转录调控分子机制的鉴定 伯基特淋巴瘤的c-myc基因。条件一直是 优化了易位与正常c-myc的分离 伯基特的几个细胞系中的基因。在约束方面的差异 已经发现了两个等位基因之间的转录因子，这些 我们会继续进行研究。金黄色葡萄球菌的鉴定与表征 体内结合的蛋白质将按照A. 将比较每个等位基因的甲基化状态。 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表达中的表达 级别对应。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。任何突变对调节性蛋白结合的影响 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c-myc激活中的作用 表情。复制c-myc易位的模型构建 将被构造并用于定义 免疫球蛋白基因座。 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 这将被定义。

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.