MOLECULAR BASIS OF ACTION OF THE PUTATIVE ONCOGENE BCL6

推定癌基因 BCL6 的分子作用基础

• 批准号: 2545410
• 负责人: VIVIAN J BARDWELL
• 金额: $ 9.62万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-30 至 2001-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2545410
• 关键词: DNA binding protein; chimeric proteins; gene expression; genetic library; intermolecular interaction; lymphoma; molecular oncology; neoplasm /cancer genetics; neoplastic cell; northern blottings; oncogenes; protein structure function; tissue /cell culture; transcription factor

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.

描述：（改编自研究者的摘要）BCL-6 是一种 DNA 结合蛋白通常在成熟 B 细胞中表达。 最终对象 拟议研究的目的是在分子水平上了解 BCL-6 如何 在正常细胞和淋巴瘤形成中发挥作用。 全部弥漫 大细胞淋巴瘤含有 BCL-6 基因、易位或点 突变，或两者兼有，可能导致 BCL-6 表达失控。 这是 BCL-6 发挥核心作用的极其有力的间接证据。 在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 作用的分子机制至关重要 鉴定 BCL-6 调节的基因并鉴定和研究 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 BCL-6 生物学的未探索领域。 一旦她了解了 BCL-6 如何 如果功能正常，她就可以开始揭示 BCL-6 在癌症中的作用。 申请人提出了两组实验。 首先，她会 鉴定受 BCL-6 直接调控的基因。 这将涉及到 生成表达功能性诱导型 BCL-6 融合的细胞系 蛋白质。 她证明了 BCL-6 与改良雌激素的融合 受体激素结合结构域使嵌合蛋白可诱导 合成抗雌激素的存在。 她将产生稳定的细胞 表达 BCL-6 融合蛋白的细胞系并使用代表性的 差异分析 (RDA) 方法用于筛选差异性 RNA 由于 BCL-6 活性增加而在这些细胞系中表达。 她 最初将限制对已知基因或具有以下基因的进一步实验 与已知基因以及显示强 BCL-6 调节的基因同源。 然后，她将通过检查这些是否会影响 BCL-6 在癌症中的作用。 BCL-6 的靶标在含有以下成分的转化细胞中受到异常调节： BCL-6 易位。 她的第二个重点是识别蛋白质 使用酵母“双杂交”遗传筛选与 BCL-6 相互作用，然后 分析它们对于 BCL-6 功能是否重要。 BCL-6 包含 蛋白质-蛋白质相互作用基序称为 POZ 结构域。 她是 对鉴定潜在的 BCL-6 伙伴蛋白特别感兴趣 通过 POZ 结构域异二聚化。 总之，拟议的研究将引导 更好地了解 BCL-6 在正常细胞中的功能以及 最终可能会解释 BCL-6 的分子病理学 淋巴瘤。