MOLECULAR GENETICS OF 11Q23 TRANSLOCATIONS

23 年 11 季度易位的分子遗传学

• 批准号: 5207593
• 负责人: ELI CANAANI
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5207593
• 关键词: acute lymphocytic leukemia; acute myelogenous leukemia; chimeric proteins; chromosome translocation; chromosomes; gene deletion mutation; genetically modified animals; human subject; immunofluorescence technique; laboratory mouse; molecular cloning; molecular oncology; neoplasm /cancer genetics; nucleic acid sequence; protein structure; site directed mutagenesis; tissue /cell culture

The ALL-1 gene located at 11q23 is rearranged in a series of reciprocal translocations with at least a dozen specific regions on a variety of chromosomes. These chromosome abnormalities are associated with acute lymphocytic leukemia and acute myeloid leukemia. Recently, we cloned the breakpoint cluster region from chromosome 11, as well as the gene spanning it, which we termed ALL-1. ALL-1 is the human homologue of Drosphilia trithorax, a gene with a critical role in regulating the Antennapedia and Bithorax homeotic gene families. In addition, we have shown that the consequence of 11q23 abnormalities is the fusion of ALL-1 to genes from the partner chromosomes and this results in production by chimeric RNAs and proteins. Three of the partner genes have now been cloned by us and others were found to share sequence homologies and/or common motifs. Here we propose to extend these findings and to clone five additional genes fused to ALL-1 in leukemias with 11q23 abnormalities, in order to determine their relationship to one another and to other genes, and to develop diagnostic tools. We propose to generate transgenic mice expressing some of the chimeric proteins to test whether the latter induce leukemia and determine which one of the reciprocal products is the oncogene. We will try to demonstrate transforming activity of the oncogene in cultured cells in order to complement the results obtained from transgenic mice, as well as to facilitate assays that could be used to examine the biological activity of in vitro-generated mutants. These studies should delineate the elements within ALL-1 and the partner proteins which play a direct role in induction of leukemia. To further investigate the mechanism by which the oncogene induces leukemia, we will generate antibodies specific to the normal and oncogenic ALL-1 proteins and use them to look for differences between the normal and oncogenic ALL-1 proteins with regard to intracellular localization, abundance, and possible aggregation with specific proteins into an active complex. In parallel, we will examine whether the presence of the oncogenic protein in cells with 11q23 abnormalities affects the abundance or the properties of the normal protein.

位于11q23的全1基因在一系列倒数中被重新排列 至少有十几个特定区域的易位 染色体。 这些染色体异常与急性有关 淋巴细胞性白血病和急性髓样白血病。 最近，我们克隆了 从11号染色体和基因的断点簇区域 跨越它，我们称其为全1。 All-1是人类的同源 interax intrithorax，一种在调节中至关重要的基因 抗抗原和bithorax同源基因家族。 此外，我们还有 表明11q23异常的结果是all-1的融合 来自伴侣染色体的基因，这导致生产 嵌合RNA和蛋白质。 现在的三个伴侣基因已经 发现我们和其他人的克隆共享序列同源和/或 共同的基础。 在这里，我们建议扩展这些发现并再加五个 为了11q23异常，在白血病中融合了all-1的基因，以便为了 确定它们彼此之间的关系以及其他基因，并与 开发诊断工具。 我们建议生成转基因小鼠 表达一些嵌合蛋白来测试后者是否 诱导白血病并确定哪种相互产品是 癌基因。 我们将尝试展示转型的活动 培养细胞中的癌基因以补充获得的结果 从转基因小鼠，以及促进可以使用的测定 检查体外生成突变体的生物学活性。 这些 研究应描述All-1和合作伙伴中的元素 在诱导白血病中发挥直接作用的蛋白质。 进一步 研究癌基因诱导白血病的机制，我们将 生成针对正常和致癌全1蛋白的抗体 并使用它们来寻找正常和致癌之间的差异 全1蛋白在细胞内定位，丰度和 可能与特定蛋白质聚集成活性复合物。 在 平行，我们将检查是否存在致癌蛋白 在11q23异常的细胞中，会影响丰度或特性 正常蛋白。