MOLECULAR BASIS OF ACTION OF THE PUTATIVE ONCOGENE BCL6

描述：(改编自研究人员的摘要)BCL-6是一种DNA 结合蛋白通常在成熟的B细胞中表达。终极目标 拟议的研究的一部分是在分子水平上了解bcl6是如何 在正常细胞和淋巴瘤的形成中起作用。所有漫反射 大细胞淋巴瘤含有bcl6基因、易位或点 突变，或两者兼而有之，很可能导致bcl6表达失控。 这是非常有力的间接证据，证明bcl-6在 在弥漫性大细胞淋巴瘤分子发病机制中的作用至 了解bcl-6作用的分子机制对于 确定bcl-6调控的基因并对其进行鉴定和研究 相互作用的蛋白质。在本提案中，申请人重点关注这些关键字 Bcl-6生物学的未知领域。一旦她了解了bcl6是如何 功能正常后，她就可以开始解开bcl6在癌症中的作用。 申请人提出了两套实验方案。首先，她会 确定受bcl6直接调控的基因。这将涉及到 构建表达功能可诱导的bcl6融合的细胞系 蛋白质。她已经证明了bcl6与一种改良的雌激素的融合 受体激素结合域使嵌合蛋白在 合成的抗雌激素的存在。她将产生稳定的细胞 表达bc1-6融合蛋白的克隆及其代表性 筛选差异RNA的差异分析(RDA)方法 在这些细胞系中的表达是bcl6活性增强的结果。她 将最初将进一步的实验限制在已知的基因或具有 与已知基因同源，以及显示强烈bcl6调控的基因。 然后，她将通过研究这些基因在癌症中的作用来解决bcl6在癌症中的作用 BCL-6的靶标在含有以下物质的转化细胞中异常调节 BCL-6易位。她的第二个重点将是识别 通过酵母“双杂交”基因筛选，与bcl-6相互作用，然后 分析它们对bcl6功能是否重要。BCL-6包含一个 蛋白质-蛋白质相互作用基序称为POZ结构域。她是 特别感兴趣的是确定潜在的bcl-6伙伴蛋白 通过POZ结构域进行异二聚化。总之，拟议的研究将导致 为了更好地了解bcl6如何在正常细胞和 最终可能导致对bcl6分子病理的解释 淋巴瘤。

DESCRIPTION: (adapted from the investigator's abstract) BCL-6 is a DNA binding protein normally expressed in mature B cells. The ultimate object of the proposed research is to understand at the molecular level how BCL-6 functions in normal cells and in the formation of lymphomas. All diffuse large cell lymphomas contain in the BCL-6 gene, translocations or point mutations, or both, which are likely to cause uncontrolled BCL-6 expression. This is extremely strong circumstantial evidence that BCL-6 plays a central role in the molecular pathogenesis of diffuse large cell lymphoma. To understand the molecular mechanisms of BCL-6 action it is critical both to identify the genes that BCL-6 regulates and to identify and study BCL-6 interacting proteins. In this proposal the applicant focuses on these key unexplored areas of the biology of BCL-6. Once she understands how BCL-6 functions normally, she can begin to unravel the role of BCL-6 in cancer. The applicant proposes two sets of experiments. In the first, she will identify genes that are directly regulated by BCL-6. This will involve generating cell lines expressing functionally inducible BCL-6 fusion proteins. She has shown that fusion of BCL-6 to a modified estrogen receptor hormone binding domain renders the chimeric protein inducible in the presence of a synthetic antiestrogen. She will generate stable cell lines expressing the BCL-6 fusion protein and use the representational difference analysis (RDA) method to screen for RNAs that are differentially expressed in these cell lines as a result of increased BCL-6 activity. She will initially restrict further experimentation to known genes or genes with homology to known genes, as well as genes that show strong BCL-6 regulation. She will then address the role of BCL-6 in cancer by examining whether these targets of BCL-6 are aberrantly regulated in transformed cells that contain BCL-6 translocations. Her second focus will be to identify proteins that interact with BCL-6, using a yeast 'two-hybrid' genetic screen, and then to analyze whether they are important for BCL-6 function. BCL-6 contains a protein-protein interaction motif called the POZ domain. She is particularly interested in identifying potential BCL-6 partner proteins that heterodimerize via the POZ domain. Together, the proposed studies will lead to a much better understanding of how BCL-6 functions in normal cells and ultimately may lead to an explanation of the molecular pathology of BCL-6 in lymphoma.