CONTROL OF C MYC TRANSCRIPTION IN HIGH GRADE LYMPHOMAS

描述：（改编自研究者摘要）申请人 建议在分子水平上研究c-myc的激活机制， 在伯基特淋巴瘤和转化淋巴瘤中， t（8;14）易位。目标是更好地了解 B细胞恶性转化的机制。 A.转录调控的分子机制 c-myc基因在B细胞中的表达。体内足迹法和体外 蛋白质-DNA结合研究将用于定位调控区域 对正常B细胞和B细胞系中c-myc表达重要 不含t（8;14）易位的细胞。的身份 体内结合的蛋白质将通过UV技术测定， 交联和免疫沉淀，然后进行杂交分析。 将对这些部位进行短暂的功能评估 在B细胞系中的转染实验。 B。转录调控的分子机制 伯基特淋巴瘤中的c-myc基因。条件已经 优化用于从正常c-myc中分离易位 几种伯基特细胞系中的基因。的约束力的差异 已经发现了两个等位基因之间的转录因子， 研究将继续进行。的鉴定和表征 体内结合的蛋白质将如A. 将比较每个等位基因的甲基化状态。 C.转录调控的分子机制 c-myc和bcl-2基因在转化型淋巴瘤中的表达及其低表达 等级对应。c-myc和bcl-2的失调， 免疫球蛋白基因座将如B中所述进行研究。 D.在调控基因中鉴定的突变对c-myc表达的影响 地区任何突变对调节蛋白结合的影响 将通过体内和体外研究来研究蛋白质。 E.免疫球蛋白位点在c-myc激活中的作用 表情复制c-myc易位的模型构建体 将被构建并用于定义 免疫球蛋白位点。 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。转录 将确定c-myc激活所需的因素 免疫球蛋白基因座的区域负责 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.