MOLECULAR BASIS OF ACTION OF THE PUTATIVE ONCOGENE BCL6

描述：（改编自研究者摘要）BCL-6是一种DNA 正常情况下在成熟B细胞中表达的结合蛋白。 终极目标 这项研究的目的是在分子水平上了解BCL-6是如何在 在正常细胞和淋巴瘤的形成中起作用。 全扩散 大细胞淋巴瘤中含有BCL-6基因，易位或点 突变，或两者，这可能导致不受控制的BCL-6表达。 这是一个非常有力的间接证据，表明BCL-6起着核心作用。 弥漫性大细胞淋巴瘤的分子发病机制中的作用。 到 了解BCL-6作用的分子机制， 确定BCL-6调控的基因，并确定和研究BCL-6 相互作用的蛋白质 在本提案中，申请人重点关注这些关键问题。 BCL-6生物学的未开发领域。 一旦她明白BCL-6 一旦BCL-6功能正常，她就可以开始揭开BCL-6在癌症中的作用。 申请人提出了两组实验。 第一，她会 确定由BCL-6直接调控的基因。 这将涉及 产生表达功能诱导型BCL-6融合蛋白的细胞系 proteins. 她已经证明BCL-6与一种修饰的雌激素的融合 受体激素结合结构域使得嵌合蛋白可诱导， 合成抗雌激素的存在 她会产生稳定的细胞 表达BCL-6融合蛋白的细胞系，并使用代表性的 差异分析（RDA）方法来筛选差异表达的RNA， 在这些细胞系中的表达是BCL-6活性增加的结果。 她 将最初限制进一步的实验，以已知的基因或基因与 与已知基因的同源性，以及显示出强BCL-6调控的基因。 然后，她将通过检查这些基因是否在癌症中发挥作用来解决BCL-6的作用。 BCL-6的靶点在转化细胞中受到异常调节， BCL-6易位。 她的第二个重点将是识别蛋白质， 与BCL-6相互作用，使用酵母“双杂交”遗传筛选，然后 分析它们对BCL-6功能是否重要。 BCL-6含有 蛋白质-蛋白质相互作用基序称为POZ结构域。 她是 特别感兴趣的是鉴定潜在的BCL-6伴侣蛋白， 通过POZ结构域异二聚化。 总之，拟议的研究将导致 更好地了解BCL-6在正常细胞中的功能， 最终可能导致对BCL-6的分子病理学的解释， 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.